PRODUCTION OF SECRETED PROTEINS BY FILAMENTOUS FUNGI

Abstract

The present invention relates to a method to improve the secretion of a protein of interest by a filamentous fungal cell comprising inducing a phenotype in the cell selected from the group consisting of a lowered ERAD, an elevated UPR that does not induce an elevated ERAD, wherein ERAD preferably is lowered. The invention further relates to the filamentous fungal cell comprising the phenotype described above. The invention also relates to polynucleotides and polypeptides whose expression can be modulated in the filamentous fungal cell to obtain the above-described phenotype.

Description

FIELD OF THE INVENTION

[0001] The invention relates to filamentous fungal cells, which display improved secretion of protein, to a method of obtaining these filamentous fungal cells and to the use of such filamentous fungal cells for production of protein.

BACKGROUND OF THE INVENTION

[0002] It is well known that filamentous fungi can be used to produce valuable compounds. Due to their glycosylation and secretion capacities, filamentous fungi are preferred hosts for secreting proteins. Secretion is a crucial step in the production of proteins and may become limiting when reaching higher production levels. High-level expression of proteins may compromise protein-folding reactions in the endoplasmic reticulum (ER), causing unfolded or aberrant proteins to accumulate. This stressful situation causes the cell to activate a variety of mechanisms. One adaptive response includes the transcriptional activation of genes encoding ER-resident chaperones and folding catalysts and protein degrading complexes that augment ER folding capacity, as well as translational attenuation to limit further accumulation of unfolded proteins in the ER (Kaufman, 1999; Mori, 2000). This signal transduction cascade is termed the unfolded protein response (UPR). Another means to deal with aberrant ER proteins is through their proteolysis via an ER-Associated Degradation (ERAD) pathway. Thus, UPR and ERAD serve one common goal, which is to decrease stress invoked by accumulation of (aberrant) proteins in the ER, either by decreasing accumulation through increasing solubility of ER localized proteins (UPR), or by increasing degradation of ER localized proteins (ERAD).

[0003] UPR and ERAD collaborate to decrease protein accumulation in the ER since it has been shown that increased UPR simultaneously results in increased ERAD (Brodsky, J. L., Werner, E. D., Dubas, M. E., Goeckeler, J. L., Kruse, K. B. and McCracken, A. A. (1999) J. Biol. Chem. 274; 3453-3460).

[0004] Recently, WO 01/72783 described a strategy to improve the protein secretion of recombinant eukaryotic cells by manipulating three genes involved in UPR (HAC1, PTC2, IRE1) in eukaryotic cells, to obtain an elevated UPR.

[0005] To improve protein secretion capacities of eukaryotic protein production strains it would be highly desirable to avail of strains that possess the capacity to translocate large amounts of a protein of interest through the secretory pathway without accumulating substantial amounts thereof in the ER.

[0006] It is an objective of the present invention to provide a method to improve protein secretion capacities of eukaryotic protein producing strains.

BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIG. 1. Disruption strategy. The basics of the disruption are depicted. Integration and subsequent removal of the disruption cassette results in removal of one of the ERAD genes.

[0008] FIG. 2. pGBFIN23. Representation of the vector that is used for over expression of genes of interest. The genes of interest were cloned as PacI-AscI fragments, placing them under control of the pepC promoter (Ppep), AmdS is used as a selectable marker.

[0009] FIG. 3. Relative glucoamylase production in Aspergillus niger strains: CBS513.88, UPR+, ERAD-, and UPR+/ERAD-. Strain CBS 513.88, UPR+, ERAD-, and UPR+/ERAD-, over expressing glucoamylase, were cultured and analyzed for glucoamylase production (example 4). The amount of extracellular glucoamylase of strain CBS 513.88 and the average of the UPR+ strains, the average of the ERAD- strains, and the average of the UPR+/ERAD- strains is depicted versus fermentation time. The highest activity of strain CBS 513.88 was set at 100%.

[0010] FIG. 4. Relative PLA2 production in Aspergillus niger strain PLA1 and three UPR+ strains. Strains PLA1, PLA-UPR1, PLA-UPR2 and PLA-UPR3, over expressing PLA2, were cultured and analyzed for PLA2 production (example 5). The amount of extracellular PLA2 is depicted, relative to strain PLA1, which was set at 100%.

[0011] FIG. 5. Relative PLA2 production in Aspergillus niger strains: PLA1 and ERAD- and ERAD- UPR+ strains derived from PLA1. Strains PLA1, PLA1-DOA, PLA1-DER, PLA1-HRD, PLA1-DOA-UPR1, PLA1-DER-UPR1, PLA1-HRD-UPR-1 and SEC61-UPR1, over expressing PLA2, were cultured and analyzed for PLA2 production (example 5). The amount of extracellular PLA2 is depicted relative to strain PLA1, which was set at 100%.

[0012] FIG. 6. Chymosin production in Aspergillus niger transformants. Strains CHY, HAC-CHY and ERP-CHY, over expressing chymosin were cultured and analyzed for chymosin production (example 6). After 6 days of fermentation the extracellular chymosin activity was determined and activities relative to strain CHY are depicted.

DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention provides, in a first aspect, a method to improve the secretion of a protein of interest by a filamentous fungal cell comprising inducing a phenotype in the cell selected from the group consisting of

[0014] (i) a lowered ERAD,

[0015] (ii) an elevated UPR that does not induce an elevated ERAD,

[0016] (iii) an elevated UPR that does not induce an elevated ERAD, wherein ERAD is lowered.

[0017] In the methods of the prior art as reflected by WO 01/72783, increased protein secretion capacities of eukaryotic protein producing strains rely on the manipulation of expression of only a single component of the secretion machinery of the cell; i.e. UPR.

[0018] Improved secretion of a protein of interest in the context of the invention means that the amount of secreted protein may be increased as compared to the parental cell the obtained cell originates from, and/or the kinetics of secretion may be elevated, and/or the quality of the protein of interest may be enhanced (e.g. higher specific activity of an enzyme by increased folding capacity of the filamentous fungal cell).

[0019] ERAD is lowered to reduce or to prevent retro-transport of the protein of interest from the ER to the cytosol, in order to reduce or to prevent its degradation. By elevating the UPR of the cell, the solubility of proteins in the ER is increased.

[0020] Changes in ERAD and/or UPR in a fungal cell may be monitored using techniques known in the art. Examples of such techniques (e.g. determining expression levels of UPR and/or ERAD related genes, pulse chase method for monitoring ERAD and biomarker assay for UPR) are described here below. A preferred assay for monitoring changes in UPR and/or ERAD is transcriptional profiling of UPR and ERAD related genes, preferably using micro arrays.

[0021] The amount of mRNA of UPR and ERAD related genes present in a cell may be monitored by transcriptional profiling (e.g. using micro arrays) and/or Northern blotting and/or real time PCR (see: Sambrook & Russell, Molecular Cloning: A Laboratory Manual, 3rd Ed., CSHL Press, Cold Spring Harbor, N.Y., 2001) and/or by quantifying the amount of corresponding protein present in a cell by Western blotting. The mRNA amount may also be monitored by DNA array analysis (Eisen, M. B. and Brown, P. O. DNA arrays for analysis of gene expression. Methods Enzymol. 1999:303:179-205).

[0022] A quantitative method may be applied to monitor ERAD. This method comprises determining the kinetics of protein secretion and degradation by the pulse chase technique as described by: Santerre Henriksen, A. L., Carlsen, M., de Bang, H. and Nielsen, J. (Kinetics of alpha-amylase secretion in Aspergillus oryzae. Biotechnol. Bioeng. 1999 Oct. 5; 65(1):76-82), and van Gemeren, I. A., Beijersbergen, A., van den Hondel C. A. and Verrips, C. T. (Expression and secretion of defined cutinase variants by Aspergillus awamori. Appl Environ. Microbiol. 1998, August; 64(8):2794-9). This pulse chase technique can be used to determine the ERAD dependent degradation kinetics of a protein of interest, when used in combination with the proteosomal inhibitor clasto-lactacystin-β-lactone (Affinity Research Products Ltd., CW8405-Z02185). Typically, the rapid degradation of proteins by ERAD is characterized by a protein half-life (t1/2) comprised between 5 and 60 minutes. t1/2 is a parameter, which can vary for each protein of interest. Preferably, t1/2 is determined for each protein. In the context of the invention, the ERAD activity is decreased when t1/2 is higher than 60 min, preferably higher than 62 min, more preferably higher than 63 min, most preferably higher than 65 min, even most preferably higher than 70 min (as described in Rabinovich, E., Kerem, A., frohlich, K. U., Diamant, N. and Bar-Nun, S. AAA-ATPase p97/Cdc48p, a cytosolic chaperone required for endoplasmic reticulum-associated protein degradation. Mol Cell Biol. 2002 January; 22(2): 626-34).

[0023] For monitoring UPR, several biomarkers are available. As a first example, it is known that the KAR2 gene, encoding the BiP protein, is induced when UPR is elevated (C. M. J. Sagt, W. H. Muller, J. Boonstra, A. J. Verkleij, and C. T. Verrips, Impaired Secretion of a Hydrophobic Cutinase by Saccharomyces cerevisiae Correlates with an Increased Association with Immunoglobulin Heavy-Chain Binding Protein (BiP) Appl. Envir. Microbiol. 1998 64: 316-324. The amount of mRNA level of KAR2 and/or of BiP protein could therefore be used as biomarker for UPR activity. Preferably, PDI or an homologue thereof is used as a biomarker (Ngiam C., Jeenes, D. J., Punt, P. J., van den Hondel, C. A. and Archer, D. B. Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger. Appl Environ Microbiol. 2000 February; 66(2):775-82.). Another preferred UPR biomarker gene is CYPB or homologue thereof (Derkx, P. M. and Madrid, S. M. The foldase CYPB is a component of the secretory pathway of Aspergillus niger and contains the endoplasmic reticulum retention signal HEEL. Mol Genet Genomics. 2001 December; 266(4): 537-45.). Another preferred biomarker gene for UPR is the spliced mRNA of hac1 mRNA or a homologue thereof (Mori. K., Ogawa, N., Kawahara, T., Yanagi, H., Yura, T. mRNA splicing-mediated C-terminal replacement of transcription factor Hac1p is required for efficient activation of the unfolded protein response. Proc Natl Acad Sci USA. 2000 Apr. 25; 97(9):4660-5, and WO 01/72783).

[0024] The fungal cell of this invention preferably is a filamentous fungal cell. A filamentous fungus is herein defined as a eukaryotic micro-organism of the subdivision Eumycota and Oomycota in filamentous form, i.e. the vegetative growth of which occurs by hyphal elongation. The filamentous fungi are characterized by a mycelia wall composed of chitin, cellulose, glucan, chitosan, mannan, and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. Filamentous fungal species include, but are not limited to, those of the genus Acremonium, Aspergillus, Aureobasidium, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Piromyces, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, and Trichoderma. Preferably, the filamentous fungal cell is selected from the group consisting of the genera Aspergillus, Trichoderma, Fusarium, Penicillium, and Acremonium. Aspergilli are mitosporic fungi characterized by an aspergillum comprised of a conidiospore stipe with no known teleomorphic states terminating in a vesicle, which in turn bears one or two layers of synchronously formed specialized cells, variously referred to as sterigmata or phialides, and asexually formed spores referred to as conidia. Known teleomorphs of Aspergillus include Eurotium, Neosartorya, and Emericella. Strains of Aspergillus and teleomorphs thereof are readily accessible to the public in a number of culture collections.

[0025] More preferably, the filamentous fungal cell is selected from the group consisting of A. nidulans, A. oryzae, A. sojae, Aspergilli of the A. niger group, Trichoderma reesei and Fusarium oxysporum. The A. niger group is herein defined according to Raper and Fennell (1965, In: The Genus Aspergillus, The Williams & Wilkins Company, Baltimore, pp 293-344) and comprises all (black) Aspergilli included in the citation. Even more preferably, the filamentous fungal cell of the present invention is selected from the group consisting of Aspergillus niger CBS 513.88, Aspergillus oryzae ATCC 20423, IFO 4177, ATCC 1011, ATCC 9576, ATCC14488-14491, ATCC 11601, ATCC12892, Aspergillus fumigatus AF293 (CBS101355), P. chrysogenum CBS 455.95, Penicillium citrinum ATCC 38065, Penicillium chrysogenum P2, Acremonium chrysogenum ATCC 36225 or ATCC 48272, Trichoderma reesei ATCC 26921 or ATCC 56765 or ATCC 26921, Aspergillus sojae ATCC11906, Chrysosporium lucknowense ATCC44006, and derivatives thereof. Most preferably, the filamentous fungal cell of the present invention is Aspergillus niger CBS 513.88. It is herein defined that A. niger CBS513.88 is a preferred parental cell to obtain the filamentous fungal cell of the present invention from and CBS513.88 is a preferred control cell in the analysis of "lowered", "elevated", "up-" or "down-regulation" of gene expression throughout the description of the present invention.

[0026] "Lowered" in the context of the present invention means at least lower as compared to the level measured in the parental cell the obtained cell originates from, the parental and obtained cell grown under the same culture conditions and analysed using the same assay conditions. Preferably, lowered means at least two times lower, more preferably at least three times lower, even more preferably at least four times lower, most preferably not detectable using Northern, or Western blotting or array analysis.

[0027] "Elevated" in the context of the present invention means at least higher as compared to the level measured in the parental cell the obtained cell originates from, the parental and obtained cell grown under the same culture conditions and analyzed using the same assay conditions. Preferably, elevated means at least two times higher, more preferably at least three times higher and most preferably at least four times higher.

[0028] The present invention also provides a method to improve the secretion of a protein of interest from a filamentous fungal cell comprising modulating the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, or a homologue thereof.

[0029] Preferably, said modulating the expression of a DNA sequence as specified above advantageously induces a phenotype in the cell selected from the group consisting of (i) a lowered ERAD, (ii) an elevated UPR that does not induce an elevated ERAD, wherein ERAD preferably is lowered.

[0030] "Modulating the expression of a DNA sequence" is defined herein that the expression of the DNA sequence may be up regulated or down regulated as compared to the expression level in the parental cell the obtained cell originates from, the parental and obtained cell grown under the same culture conditions and analyzed using the same assay conditions.

[0031] The expression level of a DNA sequence is down-regulated when the expression level of this DNA sequence in the obtained cell is lower than the expression level of the same DNA sequence in the parental cell it originates from, preferably at least two times lower, more preferably at least three times lower, even more preferably at least four times lower, most preferably not detectable.

[0032] The expression level of a DNA sequence is up regulated when its expression level is higher in the obtained cell than its expression level in the parental cell it originates from, preferably at least two times higher, more preferably at least three times higher, most preferably at least four times higher.

[0033] The modulation of the expression level of any of the above DNA sequences is preferentially monitored by transcriptional profiling using microarrays as defined previously.

[0034] According to a preferred embodiment of the invention, the expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, or a homologue thereof, is up regulated.

[0035] According to another preferred embodiment of the invention, at least one DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or a homologue thereof, is down regulated.

[0036] According to a more preferred embodiment of the invention, the expression level of at least one of the DNA sequences having the following SEQ ID NO or homologues thereof given below or a combination of at least one taken from each subgroup a), b) c), d), or e) given below, or a combination thereof is up regulated:

[0037] a) 4, 25, 34, 40,

[0038] b) 25,

[0039] c) 10, 13, 22, 25, 28, 31, 31,

[0040] d) 25,

[0041] e) 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, and/or the expression level of at least one of the DNA sequences having the following SEQ ID NO or homologues thereof, or a combination thereof is down regulated: 43, 46, 49, 52, 55, 58.

[0042] Filamentous fungal strains having:

[0043] (i) an up regulated expression level of a DNA sequence selected from the group consisting of (a) in the embodiment above, and

[0044] (ii) a down regulated expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, are particularly attractive for producing proteins rich in disulphide bridges. Proteins rich in disulphide bridges are proteins that have at least two disulphide bridges, preferably at least three and more preferably at least four. Examples of proteins rich in disulphide bridges are porcine PLA2 (seven disulphide bridges), Aspergillus phytase (five disulphide bridges) or thaumatine (eight disulphide bridges).

[0045] Filamentous fungal strains having:

[0046] (i) an up regulated expression level of a DNA sequence selected from the group consisting of (b) in the embodiment above, and

[0047] (ii) a down regulated expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, are particularly attractive for producing proteins with exposed hydrophobic patches, having a tendency to aggregate, like mutated proteins (Sagt, C. M. J., Muller, W. H., Boonstra, J., Verkleij, A. J. Verrips, C. T. Impaired secretion of a hydrophobic cutinase by Saccharomyces cerevisiae correlates with an increased association with immunoglobulin heavy-chain binding protein (BiP). Appl Environ Microbiol. 1998 January; 64(1):316-24.), or proteins unable to dimerize, or glycoproteins which are not sufficiently glycosylated (Parodi, A. J., Protein glucosylation and its role in protein folding. Annu Rev Biochem. 2000; 69:69-93. Review.).

[0048] Filamentous fungal strains having:

[0049] (i) an up regulated expression level of a DNA sequence selected from the group consisting of (c) in the embodiment above, and

[0050] (ii) a down regulated expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, are particularly attractive for producing glycoproteins, like glucoamylase or phytase.

[0051] Filamentous fungal strains having:

[0052] (i) an up regulated expression level of a DNA sequence selected from the group consisting of (d) in the embodiment above, and

[0053] (ii) a down regulated expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, are particularly attractive for producing proteins rich in proline. Proteins rich in proline are proteins that have at least 1 proline/1 kDa, preferably at least 1.5, and more preferably at least 2, like verprolin (Donnely, S. F., Pocklington, M. J., Pallotta, D., Orr, E. A proline-rich protein, verprolin, involved in cytoskeletal organization and cellular growth in the yeast Saccharomyces cerevisiae. Mol. Microbiol. 1993 November; 10(3):585-96.).

[0054] Filamentous fungal strains having:

[0055] (i) an up regulated expression level of a DNA sequence selected from the group consisting of (e) in the embodiments above, and

[0056] (ii) a down regulated expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, have good protein secretion capacities.

[0057] According to a more preferred embodiment, at least one distinct pair of genes or homologues thereof (pairs 1 to 84, table 1), or any combination of pairs is modulated (i.e. a gene is up regulated and a corresponding gene in the table is down regulated). An example of a combination of modulated pairs is: Gene pair numbers 2, 5 and 44 wherein SEQ ID NO's: 1 and 22 are up regulated and SEQ ID NO's: 46 and 55 are down regulated.

TABLE-US-00001 TABLE 1 Gene pairs to be modulated. Gene pair Up regulated Down regulated number SEQ ID NO: SEQ ID NO: 1 1 43 2 1 46 3 1 49 4 1 52 5 1 55 6 1 58 7 4 43 8 4 46 9 4 49 10 4 52 11 4 55 12 4 58 13 7 43 14 7 46 15 7 49 16 7 52 17 7 55 18 7 58 19 10 43 20 10 46 21 10 49 22 10 52 23 10 55 24 10 58 25 13 43 26 13 46 27 13 49 28 13 52 29 13 55 30 13 58 31 16 43 32 16 46 33 16 49 34 16 52 35 16 55 36 16 58 37 19 43 38 19 46 39 19 49 40 19 52 41 19 55 42 19 58 43 22 43 44 22 46 45 22 49 46 22 52 47 22 55 48 22 58 49 25 43 50 25 46 51 25 49 52 25 52 53 25 55 54 25 58 55 28 43 56 28 46 57 28 49 58 28 52 59 28 55 60 28 58 61 31 43 62 31 46 63 31 49 64 31 52 65 31 55 66 31 58 67 34 43 68 34 46 69 34 49 70 34 52 71 34 55 72 34 58 73 37 43 74 37 46 75 37 49 76 37 52 77 37 55 78 40 58 79 40 43 80 40 46 81 40 49 82 40 52 83 40 55 84 40 58

[0058] According to another more preferred embodiment: the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, or a homologue thereof, is up regulated, and

[0059] the expression level of a DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or a homologue thereof, is down regulated.

[0060] According to an even more preferred embodiment, expression of SEQ ID NO: 16 and/or SEQ ID NO: 34 is up regulated and/or expression of SEQ ID NO: 55 is down regulated. More preferably, expression of SEQ ID NO: 16 and SEQ ID NO: 34 is up regulated and/or expression of SEQ ID NO: 55 is down regulated. Even more preferably, expression of SEQ ID NO: 16 and SEQ ID NO: 34 is up regulated and expression of SEQ ID NO: 55 is down regulated.

[0061] According to another even more preferred embodiment, expression of SEQ ID NO: 16 and/or SEQ ID NO: 34 is up regulated and/or expression of SEQ ID NO: 49 is down regulated. More preferably, expression of SEQ ID NO: 16 and SEQ ID NO: 34 is up regulated and/or expression of SEQ ID NO: 49 is down regulated. Even more preferably, expression of SEQ ID NO: 16 and SEQ ID NO: 34 is up regulated and expression of SEQ ID NO: 49 is down regulated.

[0062] According to another even more preferred embodiment, expression of SEQ ID NO: 16 and/or SEQ ID NO: 34 is up regulated. More preferably, expression of SEQ ID NO: 16 and SEQ ID NO: 34 is up regulated.

[0063] According to another even more preferred embodiment, expression of SEQ ID NO: 49 is down regulated.

[0064] In addition to the above-mentioned methods, it is also possible to obtain a lowered ERAD by a specific one-way mutation of the sec61 translocation channel between ER and cytoplasm as described in WO2005/123763. Such mutation confers a phenotype wherein de novo synthesised polypeptides can enter the ER through sec61, however, retrograde transport through sec61 is impaired in the one-way mutant. In the method of this embodiment of the invention, expression of the native sec61 gene is preferably impaired, said method further comprising synthesis in the filamentous fungal cell of:

[0065] (i) a sec61 polypeptide according to SEQ ID NO: 63, and/or

[0066] (ii) a sec61 polypeptide according to SEQ ID NO: 63, wherein the amino acid at position 376 is replaced by phenylalanine, tyrosine or histidine. This specific way of lowering ERAD is preferably used in combination with above mentioned ways of lowering ERAD and/or elevated UPR.

[0067] According to a preferred embodiment of the invention, the expression level of a DNA sequence which is down regulated is lower in the obtained filamentous fungus than the expression level of the corresponding DNA sequence in the parental filamentous fungus the filamentous fungus originates from, preferably three times lower, more preferably four times lower, most preferably more than four times lower and even most preferably not detectable using northern, or western blotting or array technique.

[0068] According to another preferred embodiment of the invention, the expression level of a DNA sequence which is up regulated is higher in the obtained filamentous fungus than the expression level of the corresponding DNA sequence in the parental filamentous fungus the filamentous fungus originates from, preferably three times higher, more preferably four times higher and most preferably more than four times higher using northern, or western blotting or array technique.

[0069] The modulation of the expression level in a filamentous fungal cell of a DNA sequence as specified above may be obtained by subjecting the filamentous fungal cell to mutagenic treatment, such as recombinant genetic manipulation techniques and/or classical mutagenesis techniques, screening mutagenised cells by monitoring the expression level of said DNA sequence and, optionally, the protein secretion capacity of the filamentous fungus and identifying cells that display a modulated expression level.

[0070] Classical mutagenesis techniques comprise UV and/or chemical mutagenesis treatment commonly known in the art.

[0071] Preferably, the modulation of the expression of a DNA sequence as specified above, is achieved with recombinant genetic manipulation techniques.

[0072] The group of DNA sequences as specified above (consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61) is comprised of genomic DNA sequences. The skilled person will know that the corresponding cDNA sequences (SEQ ID NO's: 2, 5, 8, 11, 14, 17, 20, 21, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62), or homologues thereof, can be used alternatively or in combination with genomic DNA sequences in recombinant genetic techniques to achieve modulation of gene expression. Furthermore, a DNA sequence may be a synthetic nucleic acid sequence. The synthetic nucleic acid may be optimized in its codon use, preferably according to the methods described in WO2006/077258 and/or PCT/EP2007/055943.

[0073] To achieve down-regulation of a DNA sequence, said DNA sequence may be inactivated by deleting part or all of the DNA sequence or by replacing the DNA sequence by a non-functional variant thereof. The deletion and replacement may be done by gene replacement, preferably as described in EP 357 127. The specific deletion of a DNA sequence may be performed using the amdS gene as a selection marker, as described in EP 635 574.

[0074] Alternatively or in combination with other mentioned techniques, a technique based on in vivo recombination of cosmids in E. coli can be used, as described in: A rapid method for efficient gene replacement in the filamentous fungus Aspergillus nidulans (2000) Chaveroche, M-K., Ghico, J-M. and d'Enfert C; Nucleic acids Research, vol 28, no 22. This technique is applicable to other filamentous fungi like for example A. niger.

[0075] Down regulating the expression of a DNA sequence may also be achieved by using anti sense nucleic acids (see: Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger. Ngiam C, Jeenes D J, Punt P J, Van Den Hondel C A, Archer D B. Appl Environ Microbiol. 2000 February; 66(2):775-82, or Zrenner R, Willmitzer L, Sonnewald U. Analysis of the expression of potato uridinediphosphate-glucose pyrophosphorylase and its inhibition by antisense RNA. Planta. (1993);190(2):247-52.). Alternatively, down regulating expression of a DNA sequence may be achieved using RNAi techniques (see: FEMS Microb. Lett. 237 (2004): 317-324, or WO2005/05672A1, or WO2005/026356A1).

[0076] In addition to the above-mentioned techniques or as an alternative, it is also possible to obtain a lowered ERAD by inhibiting the activity of the proteins, which are involved in ERAD and encoded by a DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, or a homologue thereof. Additionally, or alternatively an ERAD-involved protein can be re-localized by means of an alternative signal sequence (Ramon de Lucas, J., Martinez O, Perez P., Isabel Lopez, M., Valenciano, S, and Laborda, F. The Aspergillus nidulans carnitine carrier encoded by the acuH gene is exclusively located in the mitochondria. FEMS Microbiol Lett. 2001 Jul. 24; 201(2):193-8.) or retention signal (Derkx, P. M. and Madrid, S. M. The foldase CYPB is a component of the secretory pathway of Aspergillus niger and contains the endoplasmic reticulum retention signal HEEL. Mol. Genet. Genomics. 2001 December; 266(4):537-45.).

[0077] Alternatively or in combination with above-mentioned techniques, inhibition of protein activity can also be obtained by UV or chemical mutagenesis (Mattern, I. E., van Noort J. M., van den Berg, P., Archer, D. B., Roberts, I. N. and van den Hondel, C. A., Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases. Mol Gen Genet. 1992 August; 234(2):332-6.) or by the use of inhibitors like the proteasomal inhibitor of Affinity (clasto-lactacystin-β-lactone, Affinity Research Products Ltd., CW8405-Z02185).

[0078] To achieve up regulation of a DNA sequence, a filamentous fungal cell may be transformed with a DNA construct comprising a DNA sequence as specified above, preferably said DNA sequence being operably linked to a promoter of a highly expressed gene. The chosen promoter may be stronger than the endogenous promoter of the DNA sequence to be over-expressed. The promoter for expression of the DNA sequence is preferably derived from a highly expressed fungal gene.

[0079] A number of preferred highly expressed fungal genes are given by way of example: the amylase, glucoamylase, alcohol dehydrogenase, xylanase, glyceraldehyde-phosphate dehydrogenase or cellobiohydrolase genes from Aspergilli or Trichoderma. Most preferred highly expressed genes for these purposes are an Aspergillus niger glucoamylase gene, an Aspergillus oryzae TAKA-amylase gene, an Aspergillus nidulans gpdA gene or a Trichoderma reesei cellobiohydrolase gene. These highly expressed genes are suitable both as target loci for integration of cloning vectors and as source of highly expressed fungal genes. The glucoamylase promoter is a preferred promoter to be used. Other preferred promoters are the promoters described in WO2006/092396 and WO2005/100573.

[0080] Up regulation may also be achieved by increasing the copy number of a DNA sequence as specified above in the eukaryotic cell, preferably by integrating into its genome copies of the DNA sequence, more preferably by targeting the integration of the DNA sequence at a highly expressed locus, for instance at a fungal glucoamylase locus.

[0081] To achieve targeted integration, an integrative cloning vector is used comprising a DNA fragment, which is homologous to a DNA sequence present in a predetermined target locus in the genome of the filamentous fungal cell, for targeting the integration of the cloning vector to this predetermined locus. In order to promote targeted integration, the cloning vector is preferably linearized prior to transformation of the filamentous fungal cell. Linearization is preferably performed such that at least one but preferably either end of the cloning vector is flanked by sequences homologous to the target locus. The length of the homologous sequences flanking the target locus is preferably at least 30 bp, preferably at least 50 bp, preferably at least 0.1 kb, even preferably at least 0.2 kb, more preferably at least 0.5 kb, even more preferably at least 1 kb, most preferably at least 2 kb. Preferably, the efficiency of targeted integration into the genome of the filamentous fungal cell, i.e. integration in a predetermined target locus, is increased by augmented homologous recombination abilities of the filamentous fungal cell. Such phenotype of the cell preferably involves a deficient ku70 gene as described in WO2005/095624. WO2005/095624 discloses a preferred method to obtain a filamentous fungal cell comprising increased efficiency of targeted integration.

[0082] Preferably, the DNA sequence in the cloning vector, which is homologous to the target locus is derived from a highly expressed locus meaning that it is derived from a gene, which is capable of high expression level in the filamentous fungal cell. A gene capable of high expression level, i.e. a highly expressed gene, is herein defined as a gene whose mRNA can make up at least 0.5% (w/w) of the total cellular mRNA, e.g. under induced conditions, or alternatively, a gene whose gene product can make up at least 1% (w/w) of the total cellular protein, or, in case of a secreted gene product, can be secreted to a level of at least 0.1 g/l (as described in EP 357 127).

[0083] To increase even more the number of copies of the DNA sequence to be over-expressed, the technique of gene conversion as described in WO98/46772 may be used.

[0084] The skilled person will appreciate the possibility that the homologous DNA sequence for targeting and the promoter sequence can coincide in one DNA fragment. The list of highly expressed genes given above is also suited as target locus.

[0085] For most filamentous fungi tested thus far it was found that they could be transformed using transformation protocols developed for Aspergillus (derived from inter alia Tilburn et al. 1983, Gene 26: 205-221). The skilled person will recognise that successful transformation of the filamentous fungal species is not limited to the use of vectors, selection marker systems, promoters and transformation protocols specifically exemplified herein. The skilled person would also understand that to obtain a filamentous fungus with both a lowered ERAD and an elevated UPR, one may use at least one of each technique described for respectively down- and up-regulating the expression of a DNA sequence in a filamentous fungus. Preferably, all the techniques performed on the filamentous fungus to obtain a recombinant filamentous fungus having both a lowered ERAD and an elevated UPR have been performed using a dominant and bi-directional selection marker, preferably an acetamidase gene, more preferably an acetamidase gene from Aspergillus nidulans or Aspergillus niger.

[0086] The transformed eukaryotic cells may subsequently be screened by monitoring the expression level of said DNA sequence as specified above by using for example Northern and/or Western blotting and/or array analysis. Optionally, the protein secretion capacity of the cell is monitored. The secretion capacity of a filamentous fungus may be monitored by measuring the amount of a protein secreted into the fermentation medium and/or the activity of a protein present in the fermentation medium after a certain fermentation period. This protein may be a marker protein or a protein of interest.

[0087] Depending on the identity of the protein of interest, the skilled person will choose a suitable detection assay. By way of example, these assay systems include but are not limited to assays based on clearing zones around colonies on solid media, as well as colorimetric, photometric, turbidimetric, viscosimetric, immunological, biological, chromatographic, and other available assays.

[0088] In a second aspect, the present invention provides a filamentous fungal cell comprising an individual feature and/or a combination of features as specified above under the first aspect. Thus, the present invention provides a filamentous fungal cell displaying a modulated expression of a DNA sequence as specified above under the first aspect.

[0089] The present invention also provides filamentous fungal cells displaying a phenotype selected from the group consisting of:

(i) a lowered ERAD, (ii) an elevated UPR that does not induce an elevated ERAD, (iii) an elevated UPR that does not induce an elevated ERAD, wherein ERAD is lowered.

[0090] In addition to modulated expression of a DNA sequence as specified under the first aspect and the phenotype described in the paragraph above, the filamentous fungal cell of the present invention may comprise by a specific one-way mutation of the sec61 translocation channel between ER and cytoplasm as described in WO2005/123763. Such mutation confers a phenotype wherein de novo synthesised polypeptides can enter the ER through sec61, however, retrograde transport through sec61 is impaired in this one-way mutant.

[0091] The filamentous fungal cell of the invention is preferably obtainable by the method as described above under the first aspect. The filamentous fungal cell of the invention may be obtained by classical genetic methods, may be a recombinant cell, or may be obtained by a combination of classical and recombinant genetic methods.

[0092] The filamentous fungal cell of the present invention preferably is a filamentous fungus as specified in the first aspect of the invention.

[0093] Optionally, the filamentous fungal cell is genetically modified to obtain a phenotype displaying lower protease expression and/or protease secretion compared to the wild-type cell in order to enhance production abilities of a polypeptide of interest. Such phenotype may be obtained by deletion and/or modification and/or inactivation of a transcriptional regulator of expression of proteases. Such a transcriptional regulator is e.g. prtT. Lowering expression of proteases by modulation of prtT may be performed by techniques described in US2004/0191864A1. Alternatively, or in combination with a phenotype displaying lower protease expression and/or protease secretion, the filamentous fungal cell displays an oxalate deficient phenotype in order to enhance the yield of production of a polypeptide of interest. An oxalate deficient phenotype may be obtained by techniques described in WO2004/070022A2. Alternatively, or in combination with a phenotype displaying lower protease expression and/or protease secretion and/or oxalate deficiency, the filamentous fungal cell displays a combination of phenotypic differences compared to the wild cell to enhance the yield of production of the polypeptide of interest. These differences may include, but are not limited to, lowered expression of glucoamylase and/or neutral alpha-amylase A and/or neutral alpha-amylase B, protease, and oxalic acid hydrolase. Said phenotypic differences displayed by the filamentous fungal cell may be obtained by genetic modification according to the techniques described in US2004/0191864A1.

[0094] In another aspect, the present invention provides a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO's: 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, or a homologue thereof. or a degenerated DNA sequence obtainable there from.

[0095] In yet another aspect, the present invention provides a polynucleotide comprising a DNA sequence encoding the polypeptide of the previous aspect. Preferably, the DNA sequence is selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, or a homologue thereof, or a degenerated DNA sequence obtainable there from.

[0096] The nucleotide sequence information as provided herein should not be so narrowly construed as to require inclusion of erroneously identified bases. The specific sequences disclosed herein can be readily used to isolate the complete gene from filamentous fungi, in particular A. niger which in turn can easily be subjected to further sequence analyses thereby identifying sequencing errors.

[0097] Unless otherwise indicated, all nucleotide sequences determined by sequencing a DNA molecule herein were determined using an automated DNA sequencer and all amino acid sequences of polypeptides encoded by DNA molecules determined herein were predicted by translation of a DNA sequence determined as above. Therefore, as is known in the art for any DNA sequence determined by this automated approach, any nucleotide sequence determined herein may contain some errors. Nucleotide sequences determined by automation are typically at least about 90% identical, more typically at least about 95% to at least about 99.9% identical to the actual nucleotide sequence of the sequenced DNA molecule. The actual sequence can be more precisely determined by other approaches including manual DNA sequencing methods well known in the art. As is also known in the art, a single insertion or deletion in a determined nucleotide sequence compared to the actual sequence will cause a frame shift in translation of the nucleotide sequence such that the predicted amino acid sequence encoded by a determined nucleotide sequence will be completely different from the amino acid sequence actually encoded by the sequenced DNA molecule, beginning at the point of such an insertion or deletion.

[0098] The person skilled in the art is capable of identifying such erroneously identified bases and knows how to correct for such errors.

[0099] In the context of the invention, a "homologue" or "homologous sequence" of a DNA sequence as specified above is defined as a DNA sequence encoding a polypeptide that displays at least one activity of the polypeptide encoded by the specified DNA sequence and has an amino acid sequence possessing a degree of identity to the amino acid sequence of the protein encoded by the specified DNA sequence of at least 50%, preferably at least 60%, more preferably at least 70%, even more preferably at least 80%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98% and most preferably at least 99%. A homologous sequence may encompass polymorphisms that may exist in cells from different populations or within a population due to natural allelic or intra-strain variation. A homologue may further be derived from a fungus other than the fungus where the specified DNA sequence originates from, or may be artificially designed and synthesized. DNA sequences related to the specified DNA sequences and obtained by degeneration of the genetic code are also part of the invention.

[0100] A "homologue" of a polypeptide is defined as a polypeptide having an amino acid sequence possessing a degree of identity to the specified amino acid sequence of at least 50%, preferably at least 60%, more preferably at least 70%, even more preferably at least 80%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98% and most preferably at least 99%, and displaying at least one activity of the polypeptide having the specified amino acid sequence.

[0101] Homologues may also encompass biologically active fragments of the full-length sequence.

[0102] For the purpose of the present invention, the degree of identity between two amino acid sequences refers to the percentage of amino acids that are identical between the two sequences. The degree of identity is determined using the BLAST algorithm, which is described in Altschul, et al., J. Mol. Biol. 215: 403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/). The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLAST program uses as defaults a word length (W) of 11, the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands. The DNA sequences of the invention may be obtained by techniques commonly known in the art. For instance by screening cDNA or genomic libraries with a suitable probe derived from a DNA sequence of the invention. It is also possible to perform PCR with suitable (degenerate) oligonucleotide primers derived from a DNA sequence of the invention. The template for such a PCR reaction may be cDNA obtained by reverse transcription of mRNA prepared from strains known or suspected to express a DNA sequence according to the invention. The PCR product may be subcloned and sequenced to ensure that the amplified sequence represents the appropriate sequence. The PCR fragment may then be used to isolate a full-length cDNA clone by a variety of known methods.

[0103] Homologues may contain only conservative substitutions of one or more amino acids of the specified amino acid sequences or substitutions, insertions or deletions of non-essential amino acids. Accordingly, a non-essential amino acid is a residue that can be altered in one of these sequences without substantially altering the biological function. For example, amino acid residues that are conserved amongst the UPR and/or ERAD proteins of the present invention, are predicted to be particularly unamenable to alteration. Furthermore, amino acids conserved among the UPR and/or ERAD proteins according to the present invention are not likely to be amenable to alteration.

[0104] The term "conservative substitution" is intended to mean that a substitution in which the amino acid residue is replaced with an amino acid residue having a similar side chain. These families are known in the art and include amino acids with basic side chains (e.g. lysine, arginine and hystidine), acidic side chains (e.g. aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagines, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0105] According to a further aspect, the present invention provides a process for the production of a protein of interest using as production organism the filamentous fungus provided by a previous aspect of this invention, the filamentous fungal cell further comprising a DNA construct comprising a DNA sequence encoding said protein of interest.

[0106] The process for the production of a protein of interest comprises culturing said filamentous fungal cell under conditions conducive to the expression of the DNA sequence encoding the protein of interest, and recovering the protein of interest, as described for example in the following references:

[0107] Li, Z. J., Shukla, V., Fordyce, A. P., Pedersen, A. G., Wenger, K. S., Marten, M. R. Fungal morphology and fragmentation behavior in a fed-batch Aspergillus oryzae fermentation at the production scale. Biotechnol Bioeng. 2000 Nov. 5; 70(3):300-12

[0108] Withers, J. M., Swift, R. J., Wiebe, M. G., Robson, G. D., Punt, P. J., van den Hondel, C. A. Optimization and stability of glucoamylase production by recombinant strains of Aspergillus niger in chemostat culture. Biotechnol Bioeng. 1998 Aug. 20; 59(4):407-18.

[0109] Amanullah, A., Christensen, L. H., Hansen, K., Nienow, A. W., Thomas, R. C. Dependence of morphology on agitation intensity in fed-batch cultures of Aspergillus oryzae and its implications for recombinant protein production. Biotechnol Bioeng. 2002 Mar. 30; 77(7):815-26.

[0110] The filamentous fungal cell of the present invention is preferably cultivated in a nutrient medium suitable for production of the polypeptide of interest. For example, the cells may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the polypeptide to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g., Bennett, J. W. and LaSure, L., eds., More Gene Manipulations in Fungi, Academic Press, CA, 1991). Suitable media are available from commercial suppliers or may be prepared using published compositions (e.g., in catalogues of the American Type Culture Collection). A suitable medium may comprise an essential cofactor for the protein of interest, e.g. flavin adenine dinucleotide (FAD). If the polypeptide is secreted into the nutrient medium, the polypeptide can be recovered directly from the medium. If the polypeptide is not secreted, it is recovered from cell lysates.

[0111] The resulting polypeptide may be isolated by methods known in the art. For example, the polypeptide may be isolated from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray drying, evaporation, or precipitation. The isolated polypeptide may then be further purified by a variety of procedures known in the art including, but not limited to, chromatography (e.g., ion exchange, affinity, hydrophobic, chromatofocusing, and size exclusion), electrophoretic procedures (e.g., preparative isoelectric focusing, differential solubility (e.g., ammonium sulfate precipitation), or extraction (see, e.g., Protein Purification, J.-C. Janson and Lars Ryden, editors, VCH Publishers, New York, 1989).

[0112] Preferably, the gene encoding the protein of interest is inserted into an expression vector, which is subsequently used to transform the filamentous fungal cell of the previous aspect. In the expression vector, the DNA sequence may be operably linked to appropriate expression signals, such as a promoter, a secretion signal sequence and a terminator, which are capable of directing the expression and secretion of the protein in the host organism.

[0113] More preferably, the gene encoding the protein of interest is operably linked to a promoter and to a secretion signal. The strategy, which can be used to express the gene encoding the protein of interest is the same as the one described for up regulating the expression of a DNA sequence: increasing copy number, targeting integration, use of a promoter of a highly expressed gene, choice of the selection marker gene, and combinations thereof. If the protein of interest is not naturally secreted, the nucleic acid encoding the protein may be modified to have a signal sequence in accordance with techniques known in the art. The secreted protein of interest may be one or more endogenous protein(s) which is (are) expressed naturally, but also may be a heterologous protein. Heterologous means that the protein is not produced under native conditions in the filamentous fungus.

[0114] The protein of interest is preferably an enzyme. Examples of enzymes which may be produced by the filamentous fungus of the invention are carbohydrases, e.g. cellulases such as endoglucanases, β-glucanases, cellobiohydrolases or β-glucosidases, hemicellulases, pectinolytic enzymes such as xylanases, xylosidases, mannanases, galactanases, galactosidases, rhamnogalacturonases, arabanases, galacturonases, lyases, or amylolytic enzymes; phosphatases such as phytases; esterases such as lipases; proteolytic enzymes; oxidoreductases such as oxidases, transferases, or isomerases.

[0115] Preferably, the filamentous fungus obtained has improved secretion capacity of the protein of interest as compared to the parental filamentous fungus it originates from.

[0116] In one embodiment, the secretion capacity of the filamentous fungal strain obtained is increased with respect to the secretion rate of the obtained strain. This rate is increased (g compound/g dry weight/hour), resulting in a decreased fermentation time, which results in a more cost effective process.

[0117] The present invention is further illustrated by the following examples.

EXAMPLES

[0118] WT1: The Aspergillus niger strain used as wild type and for internal control was already deposited under number CBS 513.88.

[0119] WT 2: This A. niger strain is a WT 1 strain comprising a deletion of the gene encoding glucoamylase (glaA). WT 2 is constructed by using the "MARKER-GENE FREE" approach as described in EP 0 635 574, wherein it is described how to delete glaA specific DNA sequences in the genome of CBS 513.88. The procedure results in a MARKER-GENE FREE ΔglaA recombinant A. niger CBS513.88 strain, possessing no foreign DNA sequences.

[0120] WT 3: This A. niger strain is a WT 2 strain comprising a deletion of the pepA gene encoding the major extracellular aspartic protease PepA. WT 3 is constructed by using the "MARKER-GENE FREE" approach as described in EP 0 635 574. The method described in this patent is used to delete pepA specific DNA sequences in the genome of CBS 513.88, as described by van den Hombergh et al. (van den Hombergh J P, Sollewijn Gelpke M D, van de Vondervoort P J, Buxton F P, Visser J. (1997)--Disruption of three acid proteases in Aspergillus niger--effects on protease spectrum, intracellular proteolysis, and degradation of target proteins--Eur J. Biochem. 247(2): 605-13). The procedure results in a MARKER-GENE FREE ΔpepA, ΔglaA recombinant A. niger CBS513.88 strain, possessing no foreign DNA sequences.

[0121] EPO1: This A. niger strain is a WT 2 strain comprising multiple copies of the A. niger epo gene coding for the proline specific endoprotease, which has been published elsewhere (WO 02/45524). EPO 1 is constructed by co-transformation of an amdS selectable marker-gene containing vector, which is designated pGBAAS-1 (constructed as described in EP 635 574) and the pGBTOPEPO-1 vector comprising the gene coding for the proline specific endoprotease as described in WO98/46772 and WO99/32617. The transformation and counterselection procedure results in a MARKER-GENE FREE EPO 1 A. niger strain containing multiple copies of the proline specific endoprotease encoding gene under control of the glucoamylase promoter.

[0122] PLA1: The heterologous porcine phospholipase A2 (PLA2) protein is selected as a model protein. It has been shown earlier that this protein is difficult to produce in A. niger in high quantities (Roberts I. N., Jeenes D. J., MacKenzie D. A., Wilkinson A. P., Sumner I. G. and Archer D. B. (1992)--Heterologous gene expression in Aspergillus niger. a glucoamylase-porcine pancreatic phospholipase A₂ fusion protein is secreted and processed to yield mature enzyme (Gene 122: 155-161). The fragment for overexpression of PLA2 is made as a fusion of propLA2 with a native glucoamylase A gene of A. niger and is prepared as described by Roberts et al. (1992). The fusion protein contains a kex1 splicing site in order to be processed in the Golgi. This glaA-pla2 fusion gene is cloned into an A. niger pGBTOP expression vector using the same techniques as described in WO 98/46772 and WO 99/32617, resulting in pGBTOPPLA-1. The PLA 1 A. niger strain is a WT 3 strain comprising multiple copies of the glucoamylase-porcine pancreatic phospholipase A₂ fusion protein encoding gene. PLA1 is constructed by co-transformation of the amdS selectable marker-gene containing vector pGBAAS-1 and the pGBTOPPLA-1 vector. The transformation and counterselection procedure results in a MARKER-GENE FREE PLA1 strain containing multiple copies of the glucoamylase-porcine pancreatic phospholipase A₂ fusion protein encoding gene under control of the glucoamylase promoter.

[0123] SEC1: The Aspergillus niger strain as strain PLA1 described above, expressing a modified sec61* translocation channel as described in WO2005/123763. This strain contains a specific one-way mutation of the sec61 translocation channel between ER and cytoplasm as described in WO2005/123763. Such mutation confers a phenotype wherein de novo synthesised polypeptides can enter the ER through sec61, however, retrograde transport through sec61 is impaired in this one-way mutant.

[0124] In these strains, using molecular biology techniques known to the skilled person (see: Sambrook & Russell, Molecular Cloning: A Laboratory Manual, 3rd Ed., CSHL Press, Cold Spring Harbor, N.Y., 2001), several genes were over expressed and others were down regulated as described below. Examples of the general design of expression vectors for gene over expression and disruption vectors for down-regulation, transformation, use of markers and selective media can be found in WO199846772, WO199932617, WO2001121779, WO2005095624, EP 635574B and WO2005100573.

A. Niger Shake Flask Fermentations

[0125] A. niger strains are precultured in 20 ml preculture medium as described in the Examples: "Aspergillus niger shake flask fermentations" section of WO 99/32617. After overnight growth, 10 ml of this culture is transferred to Fermentation Medium (FM).

[0126] Fermentation medium (FM) contains per liter: 82.5 g Glucose.1H₂O, 25 g Maldex 15 (Boom Meppel, Netherlands), 2 g Citric acid, 4.5 g NaH₂PO₄.1H₂O, 9 g KH₂PO₄, 15 g (NH₄).2SO₄, 0.02 g ZnCl₂, 0.1 g MnSO₄.1H₂O, 0.015 g CuSO₄.5H₂O, 0.015 g CoCl₂.6H₂O, 1 g MgSO₄.7H₂O, 0.1 g CaCl₂.2H₂O, 0.3 g FeSO₄.7H₂O, 30 g MES (2-[N-Morpholino]ethanesulfonic acid), pH=6.

[0127] Fermentation in FM is performed in 500 ml flasks with baffle with 100 ml fermentation broth at 34° C. and 170 rpm for the number of days indicated.

Example 1

Identification of UPR and ERAD Genes and Construction of Disruption and Overexpression Vectors

[0128] Genomic DNA of Aspergillus niger strain CBS513.88 was sequenced and analyzed (Pel et al, Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS513.88, Nature Biotechnology, Volume 25, 2, February 2007, p221-231). Sequences of all UPR & ERAD genes were identified, comprising the open reading frame (ORF) (with introns) and approximately 1000 bp 5' and 3' of the genes, are shown in sequence listings as indicated in Table 2. A number of ERAD-related genes with translated proteins annotated as described below and involved in the processes of protein secretion were named as for example derA, doaA and hrdC. In addition, UPR-related genes with translated proteins annotated as described below and involved in the processes of protein secretion were named as for example hacA, pdiA, tigA, cnxA, prpA, ostA, gptA, sstC, etc. (Table 2).

TABLE-US-00002 TABLE 2 UPR & ERAD-related genes from A. niger. Vector (overexpression Gene CDS Protein pGBFIN# or SEQ ID SEQ ID SEQ ID Gene disruption NO: NO: NO: Function (UPR/ERAD) pGBDEL#) 1 2 3 similarity to regulator of unfolded hacA pGBFINhacA protein response (UPR) Hac1 - (UPR) Saccharomyces cerevisiae 4 5 6 PDI related protein A prpA - prpA pGBFINprpA Aspergillus niger (UPR) 7 8 9 strong similarity to protein kinase Ire1 - ireA pGBFINireA Saccharomyces cerevisiae (UPR) 10 11 12 strong similarity to calcium-binding cnxK pGBFINcnxK protein precursor cnx1p - (UPR) Schizosaccharomyces pombe 13 14 15 oligosaccharyltransferase alpha ostA pGBFINostA subunit ostA - Aspergillus niger (UPR) 16 17 18 protein disulfide isomerase A pdiA - pdiA pGBFINpdiA Aspergillus niger (UPR) 19 20 21 strong similarity to ER membrane Sec61 pGBFINsec61 translocation facilitator Sec61 - (UPR) Yarrowia lipolytica 22 23 24 strong similarity to glycoprotein gptA pGBFINgptA glucosyltransferase gpt1p - (UPR) Schizosaccharomyces pombe 25 26 27 strong similarity to luminal ER-protein erdB pGBFINerdB retention receptor ERD2 - (UPR) Kluyveromyces marxianus 28 29 30 strong similarity to alpha-glucosidase modA pGBFINmodA ModA - Dictyostelium discoideum (UPR) 31 32 33 strong similarity to 80K protein H phpA pGBFINphpA precursor G19P1 - Homo sapiens (UPR) 34 35 36 strong similarity to endoplasmatic eroA pGBFINeroA reticulum oxidising protein Ero1 - (UPR) Saccharomyces cerevisiae 37 38 39 strong similarity to translation initiation sstC pGBFINsstC factor 3 47 kDa subunit stt3p - (UPR) Schizosaccharomyces pombe 40 41 42 disulfide isomerase tigA - Aspergillus tigA pGBFINtigA niger (UPR) 43 44 45 similarity to tumour suppressor protein hrdC pGBDELhrdC TSA305 from patent WO9928457-A1 - (ERAD) Homo sapiens 46 47 48 weak similarity to stress protein Herp - hrpA pGBDELhrpA Mus musculus (ERAD) 49 50 51 strong similarity to WD-repeat protein doaA pGBDELdoaA required for ubiquitin-mediated (ERAD) proteolysis Doa1 - Saccharomyces cerevisiae 52 53 54 strong similarity to protein ptcB pGBDELptcB phosphatase type 2C Ptc2 - (ERAD) Saccharomyces cerevisiae 55 56 57 strong similarity to hypothetical protein derA pGBDELderA GABA-A receptor epsilon subunit - (ERAD) Caenorhabditis elegans 58 59 60 strong similarity to alpha-mannosidase mnsA pGBDELmnsA Mns1 - Saccharomyces cerevisiae (ERAD)

[0129] Gene replacement vectors for derA, doaA and hrdC (pGBDELderA, pGBDELdoaA, pGBDELhrdC respectively) were designed according to known principles and constructed according to routine cloning procedures (see FIG. 1). In essence, these vectors comprise approximately 1000-1500 bp flanking regions of the ORFs (for SEQ ID NO. of all genes see Table 2) for homologous recombination at the predestined genomic loci. In addition, they contain the A. nidulans bi-directional amdS selection marker, in-between direct repeats. The general design of these deletion vectors were previously described in EP635574B and WO 98/46772. Additional flanking sequences for all ORF's mentioned can be found at the NCBI or EBI web servers (http://www.ncbi.nlm.nih.gov/).

[0130] DNA sequences for all UPR-related genes, such as hacA, pdiA, tigA, cnxA, prpA, ostA, gptA, sstC were cloned in expression vector pGBFIN-38 (FIG. 2) resulting in for example pGBFINostA, pGBFINgptA and pGBFINsstC, etc., and as indicated in Table 2, of which the E. coli DNA can be removed by digestion and linearization with restriction enzyme NotI, prior to transformation of the A. niger strains.

Example 2

Disruption of Genes Involved in ERAD

[0131] Disruptants of the ERAD genes were obtained by disrupting the corresponding genomic sequence using the bi-directional amdS marker. The disruption construct was designed as depicted in FIG. 1 and linear DNA of deletion vector pGBDELdoaA, pGBDELderA and pGBDELhrdC (see also Example 1) was isolated and used to transform A. niger WT1, A. niger PLA1 and A. niger EPO1 using a method earlier described (Biotechnology of Filamentous fungi: Technology and Products. (1992) Reed Publishing (USA); Chapter 6: Transformation pages 113 to 156). This linear DNA integrated into the genome at the homologous locus by gene replacement as depicted in FIG. 1, thus substituting the endogenous ERAD gene by the amdS gene. Transformants were selected on acetamide media and colony purified according to standard procedures as described in EP635574B. Spores were plated on fluoro-acetamide media to select strains, which lost the amdS marker. Growing colonies were diagnosed by PCR for integration at the doaA, derA and hrdC locus and candidate strains tested by Southern analyses for deletion of the gene. Thus, the following recombinant strains were obtained having a lowered ERAD (ERAD-, see also Table 3): WT-DOA, WT-DER, WT-HRD and PLA-DOA, PLA-DER, PLA-HRD, EPO-DOA, etc.

Example 3

Overexpression of Genes Involved in UPR

[0132] The various resulting pGBFIN overexpression vectors for UPR-related genes (such as pGBFINhacA, pGBFINpdiA pGBFINsec61, pGBFINcnxA, pGBFINerdB and pGBFINeroA pGBFINostA, pGBFINgptA and pGBFINsstC, for example) were transformed as a pool using different A. niger strains. Recipient strains in transformation were WT1 and SEC1, EPO1 and PLA1 and also a number of ERAD strains described in Example 2 using the method earlier described. The amdS gene of Aspergillus nidulans was used as selection marker and induced growth on acetamide as sole N-source as described in Kelly, J. M., and Hynes, M. J. (1985) Transformation of Aspergillus niger by the amdS gene of Aspergillus nidulans. EMBO J. 4, 475-479. The AmdS gene was placed under control of the constitutive gpdA promoter of A. nidulans. Transformants were selected on acetamide media and colony purified according to standard procedures as described in EP635574B. Growing colonies were selected for increased expression of the respective reporter genes. Subsequently, strains with increased expression levels were diagnosed by PCR for integration of one or more of the respective genes of interest and candidate strains were tested by PCR for introduction of the respective genes.

[0133] Strains mentioned as UPR+ in Table 3 (Type) were selected as representative strains for overexpression of UPR-related genes. Strains mentioned as ERAD-/UPR+ in Table 3 (Type) were selected as representative strains for overexpression of UPR-related genes in a strain background with an ERAD gene disrupted or with a modified Sec61 translocation channel as described in WO2005/123763.

TABLE-US-00003 TABLE 3 Transformation scheme for strains and constructs indicated. Transforming Parental strain plasmids Type New strain name WT1 pGBDELdoaA ERAD- WT-DOA WT1 pGBDELderA ERAD- WT-DER WT1 pGBDELhrdC ERAD- WT-HRD PLA1 pGBDELdoaA ERAD- PLA1-DOA PLA1 pGBDELderA ERAD- PLA1-DER PLA1 pGBDELhrdC ERAD- PLA1-HRD EPO1 pGBDELdoaA ERAD- EPO1-DOA EPO1 pGBDELderA ERAD- EPO1-DER EPO1 pGBDELhrdC ERAD- EPO1-HRD WT1 pGBFINostA UPR+ WT-UPR1 pGBFINgptA pGBFINsstC WT1 pGBFINostA UPR+ WT-UPR2 WT1 pGBFINgptA UPR+ WT-UPR3 pGBFINsstC PLA1 pGBFINostA UPR+ PLA-UPR1 pGBFINsstC PLA1 pGBFINgptA UPR+ PLA-UPR2 PLA1 pGBFINostA UPR+ PLA-UPR3 pGBFINgptA pGBFINsstC EPO1 pGBFINostA UPR+ EPO-UPR1 pGBFINsstC EPO1 pGBFINgptA UPR+ EPO-UPR2 WT-DOA pGBFINostA ERAD-/UPR+ WT-DOA-UPR1 pGBFINgptA pGBFINsstC WT-DER pGBFINgptA ERAD-/UPR+ WT-DER-UPR1 pGBFINsstC WT-HRD pGBFINostA ERAD-/UPR+ WT-HRD-UPR1 pGBFINgptA pGBFINsstC PLA1-DOA pGBFINprpA ERAD-/UPR+ PLA1-DOA-UPR1 pGBFINgptA pGBFINsstC PLA1-DER pGBFINpdiA ERAD-/UPR+ PLA1-DER-UPR1 pGBFINtigC PLA1-HRD pGBFINostA ERAD-/UPR+ PLA1-HRD-UPR1 pGBFINeroA EPO1-DOA pGBFINostA ERAD-/UPR+ EPO1-DOA-UPR1 pGBFINsstC EPO1-DER pGBFINostA ERAD-/UPR+ EPO1-DER-UPR1 pGBFINgptA pGBFINsstC EPO1-HRD pGBFINgptA ERAD-/UPR+ EPO1-HRD-UPR1 pGBFINsstA SEC1 pGBFINostA SEC61/UPR+ SEC1-UPR1 pGBFINgptA pGBFINsstC

[0134] This resulted in a large number of A. niger strains over expressing various combinations of UPR-related genes, disruption of ERAD genes, a modified Sec61 translocation channel and combinations thereof in different strain backgrounds, all showing increased expression of their reporter protein in a screen. The expression levels of the above-described sequences were checked by Northern analysis as described in Molecular Cloning, supra.

Example 4

Improvement of the Secretion of the Homologous Proteins Glucoamvlase and Endoprotease in the Respective ERAD and UPR A. Niger Strains of the Invention

[0135] Glycoamylase and proline specific endoprotease were used as examples of homologous secreted proteins. The endoprotease was overexpressed as described at the strains section above and the (endogenous) glucoamylase gene was expressed in the WT1 strain background.

[0136] Shake flask experiments of the UPR and ERAD strains of the EPO1 and PLA1 strains constructed in Example 2 and 3 were performed in media as described above in an incubator shaker using a 500 ml baffled shake flask. After four to six days of fermentation, samples were taken to determine either the proline specific endoprotease activity or the glucoamylase activity.

[0137] The proteolytic activity of the proline specific endoprotease is spectrophoto-metrically measured at 410 nm in time using CBZ-Gly(cine)-Pro(line)-pNA at 37° C. in a citrate/disodium phosphate buffer at pH 5. 1 U proline specific endoprotease is defined as the amount of enzyme which converts 1 μmol (micromol) CBZ-Gly(cine)-Pro(line)-pNA per min at pH 5 and 37° C. at the conditions described above.

[0138] Glucoamylase secretion was measured as the glucoamylase activity detected in the medium after 5 days of fermentation. Glucoamylase activity was measured as AGIU/ml by determining the liberation of paranitrofenol from the substrate p-nitrophenyl-a-D-glucopyranoside I. This resulted in a yellow colour, whose absorbance could be measured at 405 nm using a spectrophotometer. 1 AGIU is the quantity of enzyme, which produces 1 μmmole of glucose per minute at pH 4.3 and 60° C. from a soluble starch substrate.

[0139] The glucoamylase secretion level of all transformant strains was compared to the secretion level of Aspergillus niger WT1, which was used as a control strain. Glucoamylase production was increased in both UPR+ and in ERAD- strains compared to WT1 (Table 4 and FIG. 3). Glucoamylase production was especially increased in the ERAD-/UPR+ combination strain compared to WT1, UPR+ and ERAD- strains.

TABLE-US-00004 TABLE 4 Relative glucoamylase activities of ERAD- and UPR+ transformants New strain name Glucoamylase activity WT1 100% WT-DOA 260% WT-DER 250% WT-HRD 270% WT-UPR1 190% WT-UPR2 190% WT-UPR3 180% WT-DOA-UPR1 340% WT-DER-UPR1 290% WT-HRD-UPR1 370%

[0140] The proline-specific endoprotease secretion level of all transformant strains was compared to the secretion level of Aspergillus niger EP01, which was the recipient strain in transformation and the control strain in this experiment. Endoprotease production was increased in both UPR+ and in ERAD- strains compared to WT1 (Table 5). Also here, endoprotease production was especially increased in the ERAD-/UPR+ combination strain compared to WT1, UPR+ and ERAD- strains (Table 5). This demonstrated that the manipulation of genes involved in the UPR and/or in the ERAD lead to strains with improved protein secretion properties. Moreover, it is shown that combinatorial manipulation of down-regulation of ERAD and up-regulation of UPR has a synergetic effect on homologous protein production.

TABLE-US-00005 TABLE 5 Relative endoprotease activities of ERAD- and UPR+ transformants New strain name Endoprotease activity EPO1 100% EPO1-DOA 130% EPO1-DER 150% EPO1-HRD 140% EPO-UPR1 150% EPO-UPR2 170% EPO1-DOA-UPR1 200% EPO1-DER-UPR1 240% EPO1-HRD-UPR1 300%

Example 5

Improvement of the Secretion of a Heterologous Glucoamylase-Phospholipase A2 Fusion Protein Rich in Disulphide Bridges in the Respective ERAD and UPR A. Niger Strains of the Invention

[0141] Porcine phospholipase PLA2 was chosen as an example of a heterologous protein, which is rich in disulphide bridges. The glucoamylase-PLA2 fusion protein was over expressed under control of the glucoamylase promoter in several UPR+/ERAD- modulated strains (Table 3). The various PLA1 transformants, constructed in Example 2 and 3 and as depicted in Table 3, were fermented in media as described above in an incubator shaker using a 500 ml baffled shake flask. After four to six days of fermentation, samples were taken to determine the phospholipase activity in the fermentation medium.

[0142] To determine phospholipase PLA2 activity (PLA2) in Aspergillus niger culture broth spectrophotometrically, an artificial substrate is used: 1,2-dithiodioctanoyl phophatidylcholine (diC8, substrate). PLA2 hydrolyses the sulphide bond at the A2 position, dissociating thio-octandic acid. Thio-octandic acid reacts with 4,4 dithiopyridine (color reagent, 4-DTDP), forming 4-thiopyridone. 4-Thiopyridone is in tautomeric equilibrium with 4-mercaptopyridine, which absorbs radiation having a wavelength of 334 nm. The extinction change at that wavelength is measured. One unit is the amount of enzyme that liberates of 1 nmol thio-octandic acid from 1,2-dithiodioctanoyl phosphatidylcholine per minute at 37° C. and pH 4.0. The substrate solution is prepared by dissolving 1 g diC8 crystals per 66 ml ethanol and add 264 ml acetate buffer. The acetate buffer comprises 0.1 M Acetate buffer pH 3.85 containing 0.2% Triton-X100. The colour reagent is a 11 mM 4,4-dithiodipyridine solution. It was prepared by weighting 5.0 mg 4,4-dithiodipyridine in a 2 ml eppendorf sample cup and dissolving in 1.00 ml ethanol. 1.00 ml of milli-Q water was added.

[0143] The proline-specific PLA2 secretion level of all transformant strains was compared to the secretion level of Aspergillus niger PLA1, which was the recipient strain in transformation and the control strain in this experiment. PLA2 production was slightly increased in both UPR+ and in ERAD- strains compared to WT1 (FIGS. 4 and 5). PLA2 production was especially increased in the ERAD-/UPR+ and Sec61/UPR+ combination strains compared to WT1, UPR+ and ERAD- strains of PLA1 (FIGS. 4 and 5). This demonstrated that the manipulation of genes involved in the UPR and/or in the ERAD lead to strains with improved protein secretion properties. Moreover, it is shown that combinatorial manipulation of down-regulation of ERAD, modification of SEC61 and up-regulation of UPR has a synergetic effect on homologous protein production.

Example 6

Construction of an Aspergillus Niger Strain with Improved Secretion Capacities for a Heterologous Protein

[0144] Overexpression strains of WT3 were constructed using the methods as described in Example 3. These strains over expressed the hacA gene and pdiA and eroA genes (Table 2), respectively. The expression level of these genes was checked by Northern blot. The strains were designated HAC (hacA overexpression) and ERP (pdiA and eroA overexpression).

[0145] Calf chymosin was chosen as an example of a heterologous protein, containing disulphide bridges. This protein was over expressed under control of the glucoamylase promoter in both the WT3 strain and the obtained HAC and ERP strains using the same strategy as in Example 3, resulting in strains CHY, HAC-CHY and ERP-CHY, respectively. Transformants were selected using PCR. All transformed strains (CHY, HAC-CHY and ERP-CHY) were fermented according to the protocol as described in example 5. Chymosin concentration was determined in Milk Clotting Units (MCU) according to International Dairy Federation 157, Remcat method. The amount of extracellular chymosin activity was found to be 1.2 fold higher in the HAC-CHY strain compared to the parental strain CHY as shown in FIG. 6. The amount of extracellular chymosin activity was found to be 1.5 fold higher in the ERP-CHY strain compared to the parental strain CHY as shown in FIG. 6. Moreover if the medium was supplemented with 1 mM flavin adenine dinucleotide (FAD), the improvement in productivity for ERP-CHY was even 1.8 fold compared to the parental strain CHY (data not shown).

Sequence CWU 1

1

6311510DNAAspergillus niger 1ccacttggcc aggcctggcc cccccagctt cccccgttat gacacggtgg cctgtgttcc 60tgtgacacgg gcaagcagac gtcctccaca agctgtgtcg acctacatca ccgtcctccc 120ttgcagtgcg gttaagataa ggctcatagt aaatcgattg atccacaatt aaagatcaat 180cacctgtcac gcttgaaatg atggaagaag cattctctcc agtcgactcc ctcgccggct 240ccccgacgcc tgagttgcca ttgttgacag tgtccccggc ggacacgtcg cttgatgact 300cgtcagtaca ggcaggggag accaaggcgg aagagaagaa gcctgtgaag aagagaaagt 360catggggcca ggaattgcca gtcccgaaga ctaacttgcc cccaaggtaa gacatctata 420tccataatag actatgtatg tatgtacacg atgctaattc gacataaaag gaaacgggcc 480aagactgaag atgagaaaga gcaacgtcgt atcgagcgcg ttcttcgcaa tcgtgcggca 540gcacaaacat cacgcgagcg caagaggctc gaaatggaga agttggaaaa tgagaagatt 600cagatggaac agcaaaacca gttccttctg caacgactat cccagatgga agctgagaac 660aatcgcttaa accaacaagt cgctcaacta tctgctgagg tccggggctc ccgtggcaac 720actcccaagc ccggctcccc cgtctcagct tctcctaccc taactcctac cctatttaaa 780caagaacgcg acgaaatccc tcttgaacgg attcctttcc ccacaccctc tatcaccgac 840tactccccta ccttgaggcc ttccactctg gctgagtcct ccgacgtgac acaacatcct 900gcagcggtgt tgtgcgacct gcagtgtccg tcgctggact cgaaggagaa ggaagtgccc 960tctctctctt tgacgtcggc tcaaaccctg aacctcacgc tgccgatgat cttgcagctc 1020ctctttctga cgatgacttc caccgcctat tcaacgttga ttcacccgtt gggtcagatt 1080cttcagtcct tgaagacggg ttcgcctttg acgttctcga cggaggagat ctatcagcat 1140ttccatttga ttctatggtt gatttcgacc ccgaatctgt tggcttcgaa ggcatcgagc 1200cgccccacgg tcttccggat gagacttctc gccagacttc tagcgtgcaa cccagccttg 1260gcgcgtccac ttcgcgatgc gacgggcagg gcattgcagc tggctgttag cgagcagttt 1320cgccagggag atgcatcggc tgtcgatggt aacggagtcc aatggagctg ggagtctttg 1380ttgaccttgg cgtggacgat agacctactc gaacagccgg gacgacgcaa acgaatcttg 1440agcggtttga aatcagcgaa aactggacgg cgaagtaata ttggcaagtc tcaaaggagt 1500acacggagtt 151021026DNAAspergillus nigerCDS(1)..(1026) 2atg gaa gaa gca ttc tct cca gtc gac tcc ctc gcc ggc tcc ccg acg 48Met Glu Glu Ala Phe Ser Pro Val Asp Ser Leu Ala Gly Ser Pro Thr 1 5 10 15 cct gag ttg cca ttg ttg aca gtg tcc ccg gcg gac acg tcg ctt gat 96Pro Glu Leu Pro Leu Leu Thr Val Ser Pro Ala Asp Thr Ser Leu Asp 20 25 30 gac tcg tca gta cag gca ggg gag acc aag gcg gaa gag aag aag cct 144Asp Ser Ser Val Gln Ala Gly Glu Thr Lys Ala Glu Glu Lys Lys Pro 35 40 45 gtg aag aag aga aag tca tgg ggc cag gaa ttg cca gtc ccg aag act 192Val Lys Lys Arg Lys Ser Trp Gly Gln Glu Leu Pro Val Pro Lys Thr 50 55 60 aac ttg ccc cca agg aaa cgg gcc aag act gaa gat gag aaa gag caa 240Asn Leu Pro Pro Arg Lys Arg Ala Lys Thr Glu Asp Glu Lys Glu Gln 65 70 75 80 cgt cgt atc gag cgc gtt ctt cgc aat cgt gcg gca gca caa aca tca 288Arg Arg Ile Glu Arg Val Leu Arg Asn Arg Ala Ala Ala Gln Thr Ser 85 90 95 cgc gag cgc aag agg ctc gaa atg gag aag ttg gaa aat gag aag att 336Arg Glu Arg Lys Arg Leu Glu Met Glu Lys Leu Glu Asn Glu Lys Ile 100 105 110 cag atg gaa cag caa aac cag ttc ctt ctg caa cga cta tcc cag atg 384Gln Met Glu Gln Gln Asn Gln Phe Leu Leu Gln Arg Leu Ser Gln Met 115 120 125 gaa gct gag aac aat cgc tta aac caa caa gtc gct caa cta tct gct 432Glu Ala Glu Asn Asn Arg Leu Asn Gln Gln Val Ala Gln Leu Ser Ala 130 135 140 gag gtc cgg ggc tcc cgt ggc aac act ccc aag ccc ggc tcc ccc gtc 480Glu Val Arg Gly Ser Arg Gly Asn Thr Pro Lys Pro Gly Ser Pro Val 145 150 155 160 tca gct tct cct acc cta act cct acc cta ttt aaa caa gaa cgc gac 528Ser Ala Ser Pro Thr Leu Thr Pro Thr Leu Phe Lys Gln Glu Arg Asp 165 170 175 gaa atc cct ctt gaa cgg att cct ttc ccc aca ccc tct atc acc gac 576Glu Ile Pro Leu Glu Arg Ile Pro Phe Pro Thr Pro Ser Ile Thr Asp 180 185 190 tac tcc cct acc ttg agg cct tcc act ctg gct gag tcc tcc gac gtg 624Tyr Ser Pro Thr Leu Arg Pro Ser Thr Leu Ala Glu Ser Ser Asp Val 195 200 205 aca caa cat cct gca gtg tcc gtc gct gga ctc gaa gga gaa gga agt 672Thr Gln His Pro Ala Val Ser Val Ala Gly Leu Glu Gly Glu Gly Ser 210 215 220 gcc ctc tct ctc ttt gac gtc ggc tca aac cct gaa cct cac gct gcc 720Ala Leu Ser Leu Phe Asp Val Gly Ser Asn Pro Glu Pro His Ala Ala 225 230 235 240 gat gat ctt gca gct cct ctt tct gac gat gac ttc cac cgc cta ttc 768Asp Asp Leu Ala Ala Pro Leu Ser Asp Asp Asp Phe His Arg Leu Phe 245 250 255 aac gtt gat tca ccc gtt ggg tca gat tct tca gtc ctt gaa gac ggg 816Asn Val Asp Ser Pro Val Gly Ser Asp Ser Ser Val Leu Glu Asp Gly 260 265 270 ttc gcc ttt gac gtt ctc gac gga gga gat cta tca gca ttt cca ttt 864Phe Ala Phe Asp Val Leu Asp Gly Gly Asp Leu Ser Ala Phe Pro Phe 275 280 285 gat tct atg gtt gat ttc gac ccc gaa tct gtt ggc ttc gaa ggc atc 912Asp Ser Met Val Asp Phe Asp Pro Glu Ser Val Gly Phe Glu Gly Ile 290 295 300 gag ccg ccc cac ggt ctt ccg gat gag act tct cgc cag act tct agc 960Glu Pro Pro His Gly Leu Pro Asp Glu Thr Ser Arg Gln Thr Ser Ser 305 310 315 320 gtg caa ccc agc ctt ggc gcg tcc act tcg cga tgc gac ggg cag ggc 1008Val Gln Pro Ser Leu Gly Ala Ser Thr Ser Arg Cys Asp Gly Gln Gly 325 330 335 att gca gct ggc tgt tag 1026Ile Ala Ala Gly Cys 340 3341PRTAspergillus niger 3Met Glu Glu Ala Phe Ser Pro Val Asp Ser Leu Ala Gly Ser Pro Thr 1 5 10 15 Pro Glu Leu Pro Leu Leu Thr Val Ser Pro Ala Asp Thr Ser Leu Asp 20 25 30 Asp Ser Ser Val Gln Ala Gly Glu Thr Lys Ala Glu Glu Lys Lys Pro 35 40 45 Val Lys Lys Arg Lys Ser Trp Gly Gln Glu Leu Pro Val Pro Lys Thr 50 55 60 Asn Leu Pro Pro Arg Lys Arg Ala Lys Thr Glu Asp Glu Lys Glu Gln 65 70 75 80 Arg Arg Ile Glu Arg Val Leu Arg Asn Arg Ala Ala Ala Gln Thr Ser 85 90 95 Arg Glu Arg Lys Arg Leu Glu Met Glu Lys Leu Glu Asn Glu Lys Ile 100 105 110 Gln Met Glu Gln Gln Asn Gln Phe Leu Leu Gln Arg Leu Ser Gln Met 115 120 125 Glu Ala Glu Asn Asn Arg Leu Asn Gln Gln Val Ala Gln Leu Ser Ala 130 135 140 Glu Val Arg Gly Ser Arg Gly Asn Thr Pro Lys Pro Gly Ser Pro Val 145 150 155 160 Ser Ala Ser Pro Thr Leu Thr Pro Thr Leu Phe Lys Gln Glu Arg Asp 165 170 175 Glu Ile Pro Leu Glu Arg Ile Pro Phe Pro Thr Pro Ser Ile Thr Asp 180 185 190 Tyr Ser Pro Thr Leu Arg Pro Ser Thr Leu Ala Glu Ser Ser Asp Val 195 200 205 Thr Gln His Pro Ala Val Ser Val Ala Gly Leu Glu Gly Glu Gly Ser 210 215 220 Ala Leu Ser Leu Phe Asp Val Gly Ser Asn Pro Glu Pro His Ala Ala 225 230 235 240 Asp Asp Leu Ala Ala Pro Leu Ser Asp Asp Asp Phe His Arg Leu Phe 245 250 255 Asn Val Asp Ser Pro Val Gly Ser Asp Ser Ser Val Leu Glu Asp Gly 260 265 270 Phe Ala Phe Asp Val Leu Asp Gly Gly Asp Leu Ser Ala Phe Pro Phe 275 280 285 Asp Ser Met Val Asp Phe Asp Pro Glu Ser Val Gly Phe Glu Gly Ile 290 295 300 Glu Pro Pro His Gly Leu Pro Asp Glu Thr Ser Arg Gln Thr Ser Ser 305 310 315 320 Val Gln Pro Ser Leu Gly Ala Ser Thr Ser Arg Cys Asp Gly Gln Gly 325 330 335 Ile Ala Ala Gly Cys 340 41921DNAAspergillus niger 4gccgccattc caaatccata caattcaata ttacttctta agacatttcg cgtcacatgc 60caagagcttc aggacacctt gcttctatct acttctttct gtcctctctt ctccctctct 120tcttcctcga ttcatccccg ctgggtgatg ctttagctgc tactcttgga tcccctctcg 180catcttcctt acccgcaatc atgctgcagc ccagctctgc gttgcttttc gtcacgtcgc 240ttctggcggc gttgcccgtc aacgccgatg gattgtatac gaagaagtcc cccgtcttgc 300aggtcaacca gaagaactac gaccagctca ttgcaaactc caatcacact tcggtaagta 360cagctgtgca ggttattaca attgcctaca gacaagtcta ataagctctc ctagatcgta 420gagtaagcca tcgatcaccc tacccatcta cctcccacaa tcctaaacct ccccgctctc 480cctctagatt ctacgctccc tggtgcggcc actgccagaa cctaaagccc gcctacgaaa 540aagccgcaac taatctcgac ggcctggcca aagtcgccgc cgtcaattgc gactatgacg 600acaacaaacc cttctgcggc cgcatgggcg tccagggctt ccctaccctc aagatcgtca 660cccccggcaa gaaacccggc aagccccgcg tggaagacta caagggcgca cgaagtgcca 720aagcgattgt cgaggcagtc gtcgaccgga ttcccaacca tgtgaagcgc gcaacagaca 780aggaccttga cacttggctc gcgcaggatg aggaatcccc caaggccatc ctcttcacgg 840agaaaggcac caccagccca ctcctccgcg ccctggccat cgacttcctc ggctccatcc 900aagtcgctca agtccgcaac aaggaaaccg aagccgtcga gaaattcggc atcaccgagt 960tcccaacctt cgtcctactc ccaggaggcg gccaagaccc catcgtctac gacggcgaac 1020tgaagaagaa gcccatggtc gaattcctca gccaagccgc tgctcctaac ccggatcctg 1080ctcccaaggg ctcgaccgcg ccccgcgata acaacaagaa gaaatccacc gaaccttctc 1140cagactccaa gattgtctcg gacgaggcca aacccgccag tgtgcccatt ccggctcccc 1200ccattggtac cctgcccact gcggaagccc tcgaggctgc ttgtctgatg ccgaaatccg 1260gtacctgtgt gctggctctc ctccctgaac cgagtgagcc ggacgcagag ctcccggctc 1320cggccaagga cgccctcctc agtctcgctg agatctcgca caagcacgca gtccgtaaga 1380gcaagctctt cccgttctac agtgtcccgg ctatcaatag cggagctaag accctccgcg 1440ctgggcttgg tctgcctgag gataactcgg tggagatcgt tgctgtgaat ggacgccgtg 1500gctggtggcg ccggtatgac tcggttgagg gcgcagagta cggccaggag cgtgtcgagg 1560cttggattga tgcgatcagg ctgggtgagg gtgagaagca gaagttgcct gatggcgttg 1620tcgttgaaga ggtagttgag gagaaggtcg aagagaaggt cgaggaagtg gttgaagaac 1680ccgtcgagga gaagccggcg gtcgaccacg acgaattgta aaacatatgg tccgtatgga 1740gtgcatgaat ttgtttatta gcacaggtgt ttatcaggtc aaataagtac tactagctgg 1800tttcccatat cgagtatcaa aagcatacat atcatctact gtcagctact tcaattccac 1860taatcgggat gaacttgtat tggaacactc atgtagaaat aagctctcta aagattcaat 1920t 192151395DNAAspergillus nigerCDS(1)..(1395) 5atg ctg cag ccc agc tct gcg ttg ctt ttc gtc acg tcg ctt ctg gcg 48Met Leu Gln Pro Ser Ser Ala Leu Leu Phe Val Thr Ser Leu Leu Ala 1 5 10 15 gcg ttg ccc gtc aac gcc gat gga ttg tat acg aag aag tcc ccc gtc 96Ala Leu Pro Val Asn Ala Asp Gly Leu Tyr Thr Lys Lys Ser Pro Val 20 25 30 ttg cag gtc aac cag aag aac tac gac cag ctc att gca aac tcc aat 144Leu Gln Val Asn Gln Lys Asn Tyr Asp Gln Leu Ile Ala Asn Ser Asn 35 40 45 cac act tcg atc gta gaa ttc tac gct ccc tgg tgc ggc cac tgc cag 192His Thr Ser Ile Val Glu Phe Tyr Ala Pro Trp Cys Gly His Cys Gln 50 55 60 aac cta aag ccc gcc tac gaa aaa gcc gca act aat ctc gac ggc ctg 240Asn Leu Lys Pro Ala Tyr Glu Lys Ala Ala Thr Asn Leu Asp Gly Leu 65 70 75 80 gcc aaa gtc gcc gcc gtc aat tgc gac tat gac gac aac aaa ccc ttc 288Ala Lys Val Ala Ala Val Asn Cys Asp Tyr Asp Asp Asn Lys Pro Phe 85 90 95 tgc ggc cgc atg ggc gtc cag ggc ttc cct acc ctc aag atc gtc acc 336Cys Gly Arg Met Gly Val Gln Gly Phe Pro Thr Leu Lys Ile Val Thr 100 105 110 ccc ggc aag aaa ccc ggc aag ccc cgc gtg gaa gac tac aag ggc gca 384Pro Gly Lys Lys Pro Gly Lys Pro Arg Val Glu Asp Tyr Lys Gly Ala 115 120 125 cga agt gcc aaa gcg att gtc gag gca gtc gtc gac cgg att ccc aac 432Arg Ser Ala Lys Ala Ile Val Glu Ala Val Val Asp Arg Ile Pro Asn 130 135 140 cat gtg aag cgc gca aca gac aag gac ctt gac act tgg ctc gcg cag 480His Val Lys Arg Ala Thr Asp Lys Asp Leu Asp Thr Trp Leu Ala Gln 145 150 155 160 gat gag gaa tcc ccc aag gcc atc ctc ttc acg gag aaa ggc acc acc 528Asp Glu Glu Ser Pro Lys Ala Ile Leu Phe Thr Glu Lys Gly Thr Thr 165 170 175 agc cca ctc ctc cgc gcc ctg gcc atc gac ttc ctc ggc tcc atc caa 576Ser Pro Leu Leu Arg Ala Leu Ala Ile Asp Phe Leu Gly Ser Ile Gln 180 185 190 gtc gct caa gtc cgc aac aag gaa acc gaa gcc gtc gag aaa ttc ggc 624Val Ala Gln Val Arg Asn Lys Glu Thr Glu Ala Val Glu Lys Phe Gly 195 200 205 atc acc gag ttc cca acc ttc gtc cta ctc cca gga ggc ggc caa gac 672Ile Thr Glu Phe Pro Thr Phe Val Leu Leu Pro Gly Gly Gly Gln Asp 210 215 220 ccc atc gtc tac gac ggc gaa ctg aag aag aag ccc atg gtc gaa ttc 720Pro Ile Val Tyr Asp Gly Glu Leu Lys Lys Lys Pro Met Val Glu Phe 225 230 235 240 ctc agc caa gcc gct gct cct aac ccg gat cct gct ccc aag ggc tcg 768Leu Ser Gln Ala Ala Ala Pro Asn Pro Asp Pro Ala Pro Lys Gly Ser 245 250 255 acc gcg ccc cgc gat aac aac aag aag aaa tcc acc gaa cct tct cca 816Thr Ala Pro Arg Asp Asn Asn Lys Lys Lys Ser Thr Glu Pro Ser Pro 260 265 270 gac tcc aag att gtc tcg gac gag gcc aaa ccc gcc agt gtg ccc att 864Asp Ser Lys Ile Val Ser Asp Glu Ala Lys Pro Ala Ser Val Pro Ile 275 280 285 ccg gct ccc ccc att ggt acc ctg ccc act gcg gaa gcc ctc gag gct 912Pro Ala Pro Pro Ile Gly Thr Leu Pro Thr Ala Glu Ala Leu Glu Ala 290 295 300 gct tgt ctg atg ccg aaa tcc ggt acc tgt gtg ctg gct ctc ctc cct 960Ala Cys Leu Met Pro Lys Ser Gly Thr Cys Val Leu Ala Leu Leu Pro 305 310 315 320 gaa ccg agt gag ccg gac gca gag ctc ccg gct ccg gcc aag gac gcc 1008Glu Pro Ser Glu Pro Asp Ala Glu Leu Pro Ala Pro Ala Lys Asp Ala 325 330 335 ctc ctc agt ctc gct gag atc tcg cac aag cac gca gtc cgt aag agc 1056Leu Leu Ser Leu Ala Glu Ile Ser His Lys His Ala Val Arg Lys Ser 340 345 350 aag ctc ttc ccg ttc tac agt gtc ccg gct atc aat agc gga gct aag 1104Lys Leu Phe Pro Phe Tyr Ser Val Pro Ala Ile Asn Ser Gly Ala Lys 355 360 365 acc ctc cgc gct ggg ctt ggt ctg cct gag gat aac tcg gtg gag atc 1152Thr Leu Arg Ala Gly Leu Gly Leu Pro Glu Asp Asn Ser Val Glu Ile 370 375 380 gtt gct gtg aat gga cgc cgt ggc tgg tgg cgc cgg tat gac tcg gtt 1200Val Ala Val Asn Gly Arg Arg Gly Trp Trp Arg Arg Tyr Asp Ser Val 385 390 395 400 gag ggc gca gag tac ggc cag gag cgt gtc gag gct tgg att gat gcg 1248Glu Gly Ala Glu Tyr Gly Gln Glu Arg Val Glu Ala Trp Ile Asp Ala 405 410 415 atc agg ctg ggt gag ggt gag aag cag aag ttg cct gat ggc gtt gtc 1296Ile Arg Leu Gly Glu Gly Glu Lys Gln Lys Leu Pro Asp Gly Val Val 420 425 430 gtt gaa gag gta gtt gag gag aag gtc gaa gag aag gtc gag gaa gtg 1344Val Glu Glu Val Val Glu Glu Lys Val Glu Glu Lys Val Glu Glu Val 435 440 445 gtt gaa gaa ccc gtc gag gag aag ccg gcg gtc gac cac gac gaa ttg 1392Val Glu Glu Pro Val Glu Glu Lys Pro Ala Val Asp His Asp Glu Leu 450 455 460 taa 13956464PRTAspergillus niger 6Met Leu Gln Pro Ser Ser Ala Leu Leu Phe Val Thr Ser Leu Leu Ala 1 5 10 15 Ala Leu Pro Val Asn Ala Asp Gly Leu Tyr Thr Lys Lys Ser Pro Val 20 25 30 Leu Gln Val Asn Gln Lys Asn Tyr Asp Gln Leu Ile Ala Asn Ser Asn 35 40 45 His Thr Ser Ile Val Glu Phe Tyr Ala Pro Trp Cys Gly His Cys Gln 50 55 60 Asn Leu Lys Pro Ala Tyr Glu Lys Ala Ala Thr Asn Leu Asp Gly Leu 65 70 75 80 Ala Lys Val Ala Ala Val Asn Cys Asp Tyr Asp Asp Asn Lys Pro Phe 85 90 95 Cys Gly Arg Met Gly Val Gln Gly Phe Pro Thr Leu Lys Ile Val Thr 100 105 110 Pro Gly

Lys Lys Pro Gly Lys Pro Arg Val Glu Asp Tyr Lys Gly Ala 115 120 125 Arg Ser Ala Lys Ala Ile Val Glu Ala Val Val Asp Arg Ile Pro Asn 130 135 140 His Val Lys Arg Ala Thr Asp Lys Asp Leu Asp Thr Trp Leu Ala Gln 145 150 155 160 Asp Glu Glu Ser Pro Lys Ala Ile Leu Phe Thr Glu Lys Gly Thr Thr 165 170 175 Ser Pro Leu Leu Arg Ala Leu Ala Ile Asp Phe Leu Gly Ser Ile Gln 180 185 190 Val Ala Gln Val Arg Asn Lys Glu Thr Glu Ala Val Glu Lys Phe Gly 195 200 205 Ile Thr Glu Phe Pro Thr Phe Val Leu Leu Pro Gly Gly Gly Gln Asp 210 215 220 Pro Ile Val Tyr Asp Gly Glu Leu Lys Lys Lys Pro Met Val Glu Phe 225 230 235 240 Leu Ser Gln Ala Ala Ala Pro Asn Pro Asp Pro Ala Pro Lys Gly Ser 245 250 255 Thr Ala Pro Arg Asp Asn Asn Lys Lys Lys Ser Thr Glu Pro Ser Pro 260 265 270 Asp Ser Lys Ile Val Ser Asp Glu Ala Lys Pro Ala Ser Val Pro Ile 275 280 285 Pro Ala Pro Pro Ile Gly Thr Leu Pro Thr Ala Glu Ala Leu Glu Ala 290 295 300 Ala Cys Leu Met Pro Lys Ser Gly Thr Cys Val Leu Ala Leu Leu Pro 305 310 315 320 Glu Pro Ser Glu Pro Asp Ala Glu Leu Pro Ala Pro Ala Lys Asp Ala 325 330 335 Leu Leu Ser Leu Ala Glu Ile Ser His Lys His Ala Val Arg Lys Ser 340 345 350 Lys Leu Phe Pro Phe Tyr Ser Val Pro Ala Ile Asn Ser Gly Ala Lys 355 360 365 Thr Leu Arg Ala Gly Leu Gly Leu Pro Glu Asp Asn Ser Val Glu Ile 370 375 380 Val Ala Val Asn Gly Arg Arg Gly Trp Trp Arg Arg Tyr Asp Ser Val 385 390 395 400 Glu Gly Ala Glu Tyr Gly Gln Glu Arg Val Glu Ala Trp Ile Asp Ala 405 410 415 Ile Arg Leu Gly Glu Gly Glu Lys Gln Lys Leu Pro Asp Gly Val Val 420 425 430 Val Glu Glu Val Val Glu Glu Lys Val Glu Glu Lys Val Glu Glu Val 435 440 445 Val Glu Glu Pro Val Glu Glu Lys Pro Ala Val Asp His Asp Glu Leu 450 455 460 73891DNAAspergillus niger 7atccaattca tccattctat tccatcctat tcatccgctc aatgctgccc ttcaattgcc 60catctgctct cgattcttct tccttctctt ttggttcctc ccacgcggat tttaccaact 120gatgacacgc cccgtgccat cggatccctc actctccagc ttctctctcc atctggccca 180ctgtattgga gtccctcagc atgcggtggc ggctgcctgg cgcccggtcg acccttcctg 240ccagtgtcgc actcctcctg ctccccgttc ttgttgctcc gcagcagtgg catgaacatc 300aacatgagct ctcctccacc gtttccgtcc ctctccgacc gactggtttc acctccggcg 360tcgatacccc tccctctttc gacgtgaaat ccaacgatgc gagcgcccta gcaaccctgg 420ctctggccgg ctctggccgc gccgttcgag cccctcctgc ccaagccagc agctctaccg 480ctggcctggc tccgcagctt cacgcgcggt ccctgcagga ctgggaggtt gaggactttg 540tcctgctggc gaccgtcgac ggttccattc acgcacgcga ccgcaagacc ggtgccgctc 600gttgggccct cgaggtcccg agcagcccta tggtcgaaag cctctaccac cgagccaatc 660gctccagctt cgaccgtgcc caaccagagg acgactttat ctggatcgtc gagccgagtc 720agggcggaag cctctacatc tacagctcgg ggccagaggc aggcctccag aaattgggat 780tgactgtgaa ggaacttgtt gacgaaacgc cttactcggg gactgacccg gccgttactt 840atacggcacg aaaggaaacg acgctgtata ccatcgatgc tcgcaccgga aacattctgc 900gggtgtttag ctctagaggt cccatttcgt caggtcagga atgtcgaaag gttgatggtc 960tggatgtgga tatggaagaa tgcgaatccc cttcgggtac tctagtcctt ggtcgtgtcg 1020aatacacggt agccatccag aacaccgaaa ccggtgatcc aatctgcact ctcaagtact 1080cggagtggac ggccaacaac cgggatatgg acctccagag ccagtacctc cgcacgatgg 1140atcaaagcca tatttacagc atgcatgatg gtgtagtctt aggcttcgat cattcacgga 1200tggaccggcc acggtacacc cagcgattct cgagtccggt ggtccgcgtc ttcgatgttg 1260ctcgtccggt cagcgccgac tcatctaacg accctactcc acttattcta ctctcgcagc 1320ctctacagcc tcctgacccc gactacggta cgcttgacga tcgtgatgaa agagtattca 1380ttgattacac cgagggtggt ggttggtatg ccatgtcgga ggccacctac ccgcttgtca 1440ccgggagagc caagatggct caatgctacg aaaaagatta cctccgccat ggtcaacccc 1500taacaagtct gaccccgagt cagcaacaag atgcactagc aggagtccat tctttgaacg 1560gcccacgcgt cgtccgccgt cacatcccca gcatttctgg cccctcgtca gccgatatgt 1620ccaatgacac gcctcgggag ttgatctata gctcatcgga cttggcactg cctccggctc 1680tacgccacag caccattata cggaagggct gggacaatgc cattgatatt tttgtgacgc 1740tcttgcttct gtttttcggc accttcatct ggttcaattc tcatcacatt caggagcttg 1800ctaagcagaa gctggatctg aaaaatatca tggcctcgta cggacagccg cccatgtcta 1860ccccctcaac tccaatcgtg gaagcccctc atttgaaacg cgaggctagc cctaatcgca 1920tggcgaatct gactgtcgac atgaatgttt caggagagca gccgcagggt ggtgactcga 1980cgccaaggcc caagaaatcc cagaactctc ttgcgcccga cacaactcca cgcgtacgca 2040tccgggaacc gtctcaaggc ccagatggcg atgacgatgt ggacgagctc aatctacaag 2100acggtgaaaa gcctaagaag aaggctcgcc gcggtcgtcg tggtggcaag aatcataggc 2160ggggcaagaa gcccaatagc gacagcgaat ccagggaccc ggccgatcgc gttgttgatg 2220aagtgaacaa gcttcaacct cagcctcgct tggaacccga tgtacagctg gcccggacgg 2280tgtcgcatga gatcatggaa atggatggcg ttctccagat cggccgtctt agggtgttca 2340ctgacgtggt cctgggacac ggcagccacg ggaccgtggt gtatcggggc tcgttcgatg 2400gacgcgacgt ggctgtcaag cgcatgctgg tagaattcta tgatattgca tcccatgaag 2460tgggcctgtt gcaagaaagt gatgaccatg gcaatgtgat ccggtactac tgccgagagc 2520aggctgctgg tttcctctac attgctttgg agctctgccc ggcctctttg caggatgtgg 2580ttgaacgtcc atcagatttc ccgcagttag tccagggcgg cttggacctg ccggacgttc 2640tgcgccagat tgtggcaggt gttcgctatc ttcattctct taagattgtg caccgcgatc 2700tgaagccaca gaacatcttg gtggcgatgc ctcgcgggcg tactggttca cgctccctgc 2760ggttgctgat ctcggatttc ggcttgtgta agaagctcga agacaaccag agctccttcc 2820gcgcaactac ggcacatgcc gcgggtacct caggctggcg agcccctgaa ttgctggtag 2880acgacgacat gagcccggct atgcagggta gcgagtccca acacaccgaa tcatcagaac 2940cagctgtggt ggatcctcaa accaaccggc gggctactcg agctatcgac atcttctctt 3000tgggctgcgt cttttattac gttctgacgc gggggtgcca tccttttgac aagaatggca 3060agtttatgcg cgaggccaac attgtcaagg gcaaccacaa cctcgatgag ctgcagcgtc 3120tgggcgacta tgcctacgag gctgaagatc taatccagtc catgttgtcg cttgatcctc 3180gacgacggta agtcgatgct cattacgtgc catgcatagt actaactttt ctagacccga 3240tgcgagcgct gtgttgacgc acccgttctt ttggcctcca tctgaccgtc ttagcttcct 3300ctgcgatgtc tcggatcact ttgaatttga accgcgggat cctccttcgg acgccctttt 3360gtgtctcgag tcggtcgctc cacgagtgat gggcccggac atggatttcc tgcgactact 3420gccacgggac tttaaggata atctcggcaa gcagcgtaag tacacgggat cgaagatgtt 3480agatttgctg cgagccctcc ggaacaagcg caaccattac aacgacatgc cggagcatct 3540caaggcacac atcggcgggt tgcccgaggg gtatcttaat ttttggactg tgcgattccc 3600cagtcttctc atgagctgcc actccgtcat tgtggagttg cgtttgacgc ggtccgaccg 3660tttcaagcgc tacttcacgg cgactgacta ggtggtgttc acccacgtag acagtcattt 3720acttgtatac atgcatatct agatgacata tgtcacaatc aataagttat acgagtctta 3780cttatcattc tatcaatggg aattcatgct gcagagtcgt ccggtagtgt gggcggggta 3840gtcacgtgtc tagtctagtg accggggaag ccctcagcga tgagtcatgg a 389183444DNAAspergillus nigerCDS(1)..(3444) 8atg cgg tgg cgg ctg cct ggc gcc cgg tcg acc ctt cct gcc agt gtc 48Met Arg Trp Arg Leu Pro Gly Ala Arg Ser Thr Leu Pro Ala Ser Val 1 5 10 15 gca ctc ctc ctg ctc ccc gtt ctt gtt gct ccg cag cag tgg cat gaa 96Ala Leu Leu Leu Leu Pro Val Leu Val Ala Pro Gln Gln Trp His Glu 20 25 30 cat caa cat gag ctc tcc tcc acc gtt tcc gtc cct ctc cga ccg act 144His Gln His Glu Leu Ser Ser Thr Val Ser Val Pro Leu Arg Pro Thr 35 40 45 ggt ttc acc tcc ggc gtc gat acc cct ccc tct ttc gac gtg aaa tcc 192Gly Phe Thr Ser Gly Val Asp Thr Pro Pro Ser Phe Asp Val Lys Ser 50 55 60 aac gat gcg agc gcc cta gca acc ctg gct ctg gcc ggc tct ggc cgc 240Asn Asp Ala Ser Ala Leu Ala Thr Leu Ala Leu Ala Gly Ser Gly Arg 65 70 75 80 gcc gtt cga gcc cct cct gcc caa gcc agc agc tct acc gct ggc ctg 288Ala Val Arg Ala Pro Pro Ala Gln Ala Ser Ser Ser Thr Ala Gly Leu 85 90 95 gct ccg cag ctt cac gcg cgg tcc ctg cag gac tgg gag gtt gag gac 336Ala Pro Gln Leu His Ala Arg Ser Leu Gln Asp Trp Glu Val Glu Asp 100 105 110 ttt gtc ctg ctg gcg acc gtc gac ggt tcc att cac gca cgc gac cgc 384Phe Val Leu Leu Ala Thr Val Asp Gly Ser Ile His Ala Arg Asp Arg 115 120 125 aag acc ggt gcc gct cgt tgg gcc ctc gag gtc ccg agc agc cct atg 432Lys Thr Gly Ala Ala Arg Trp Ala Leu Glu Val Pro Ser Ser Pro Met 130 135 140 gtc gaa agc ctc tac cac cga gcc aat cgc tcc agc ttc gac cgt gcc 480Val Glu Ser Leu Tyr His Arg Ala Asn Arg Ser Ser Phe Asp Arg Ala 145 150 155 160 caa cca gag gac gac ttt atc tgg atc gtc gag ccg agt cag ggc gga 528Gln Pro Glu Asp Asp Phe Ile Trp Ile Val Glu Pro Ser Gln Gly Gly 165 170 175 agc ctc tac atc tac agc tcg ggg cca gag gca ggc ctc cag aaa ttg 576Ser Leu Tyr Ile Tyr Ser Ser Gly Pro Glu Ala Gly Leu Gln Lys Leu 180 185 190 gga ttg act gtg aag gaa ctt gtt gac gaa acg cct tac tcg ggg act 624Gly Leu Thr Val Lys Glu Leu Val Asp Glu Thr Pro Tyr Ser Gly Thr 195 200 205 gac ccg gcc gtt act tat acg gca cga aag gaa acg acg ctg tat acc 672Asp Pro Ala Val Thr Tyr Thr Ala Arg Lys Glu Thr Thr Leu Tyr Thr 210 215 220 atc gat gct cgc acc gga aac att ctg cgg gtg ttt agc tct aga ggt 720Ile Asp Ala Arg Thr Gly Asn Ile Leu Arg Val Phe Ser Ser Arg Gly 225 230 235 240 ccc att tcg tca ggt cag gaa tgt cga aag gtt gat ggt ctg gat gtg 768Pro Ile Ser Ser Gly Gln Glu Cys Arg Lys Val Asp Gly Leu Asp Val 245 250 255 gat atg gaa gaa tgc gaa tcc cct tcg ggt act cta gtc ctt ggt cgt 816Asp Met Glu Glu Cys Glu Ser Pro Ser Gly Thr Leu Val Leu Gly Arg 260 265 270 gtc gaa tac acg gta gcc atc cag aac acc gaa acc ggt gat cca atc 864Val Glu Tyr Thr Val Ala Ile Gln Asn Thr Glu Thr Gly Asp Pro Ile 275 280 285 tgc act ctc aag tac tcg gag tgg acg gcc aac aac cgg gat atg gac 912Cys Thr Leu Lys Tyr Ser Glu Trp Thr Ala Asn Asn Arg Asp Met Asp 290 295 300 ctc cag agc cag tac ctc cgc acg atg gat caa agc cat att tac agc 960Leu Gln Ser Gln Tyr Leu Arg Thr Met Asp Gln Ser His Ile Tyr Ser 305 310 315 320 atg cat gat ggt gta gtc tta ggc ttc gat cat tca cgg atg gac cgg 1008Met His Asp Gly Val Val Leu Gly Phe Asp His Ser Arg Met Asp Arg 325 330 335 cca cgg tac acc cag cga ttc tcg agt ccg gtg gtc cgc gtc ttc gat 1056Pro Arg Tyr Thr Gln Arg Phe Ser Ser Pro Val Val Arg Val Phe Asp 340 345 350 gtt gct cgt ccg gtc agc gcc gac tca tct aac gac cct act cca ctt 1104Val Ala Arg Pro Val Ser Ala Asp Ser Ser Asn Asp Pro Thr Pro Leu 355 360 365 att cta ctc tcg cag cct cta cag cct cct gac ccc gac tac ggt acg 1152Ile Leu Leu Ser Gln Pro Leu Gln Pro Pro Asp Pro Asp Tyr Gly Thr 370 375 380 ctt gac gat cgt gat gaa aga gta ttc att gat tac acc gag ggt ggt 1200Leu Asp Asp Arg Asp Glu Arg Val Phe Ile Asp Tyr Thr Glu Gly Gly 385 390 395 400 ggt tgg tat gcc atg tcg gag gcc acc tac ccg ctt gtc acc ggg aga 1248Gly Trp Tyr Ala Met Ser Glu Ala Thr Tyr Pro Leu Val Thr Gly Arg 405 410 415 gcc aag atg gct caa tgc tac gaa aaa gat tac ctc cgc cat ggt caa 1296Ala Lys Met Ala Gln Cys Tyr Glu Lys Asp Tyr Leu Arg His Gly Gln 420 425 430 ccc cta aca agt ctg acc ccg agt cag caa caa gat gca cta gca gga 1344Pro Leu Thr Ser Leu Thr Pro Ser Gln Gln Gln Asp Ala Leu Ala Gly 435 440 445 gtc cat tct ttg aac ggc cca cgc gtc gtc cgc cgt cac atc ccc agc 1392Val His Ser Leu Asn Gly Pro Arg Val Val Arg Arg His Ile Pro Ser 450 455 460 att tct ggc ccc tcg tca gcc gat atg tcc aat gac acg cct cgg gag 1440Ile Ser Gly Pro Ser Ser Ala Asp Met Ser Asn Asp Thr Pro Arg Glu 465 470 475 480 ttg atc tat agc tca tcg gac ttg gca ctg cct ccg gct cta cgc cac 1488Leu Ile Tyr Ser Ser Ser Asp Leu Ala Leu Pro Pro Ala Leu Arg His 485 490 495 agc acc att ata cgg aag ggc tgg gac aat gcc att gat att ttt gtg 1536Ser Thr Ile Ile Arg Lys Gly Trp Asp Asn Ala Ile Asp Ile Phe Val 500 505 510 acg ctc ttg ctt ctg ttt ttc ggc acc ttc atc tgg ttc aat tct cat 1584Thr Leu Leu Leu Leu Phe Phe Gly Thr Phe Ile Trp Phe Asn Ser His 515 520 525 cac att cag gag ctt gct aag cag aag ctg gat ctg aaa aat atc atg 1632His Ile Gln Glu Leu Ala Lys Gln Lys Leu Asp Leu Lys Asn Ile Met 530 535 540 gcc tcg tac gga cag ccg ccc atg tct acc ccc tca act cca atc gtg 1680Ala Ser Tyr Gly Gln Pro Pro Met Ser Thr Pro Ser Thr Pro Ile Val 545 550 555 560 gaa gcc cct cat ttg aaa cgc gag gct agc cct aat cgc atg gcg aat 1728Glu Ala Pro His Leu Lys Arg Glu Ala Ser Pro Asn Arg Met Ala Asn 565 570 575 ctg act gtc gac atg aat gtt tca gga gag cag ccg cag ggt ggt gac 1776Leu Thr Val Asp Met Asn Val Ser Gly Glu Gln Pro Gln Gly Gly Asp 580 585 590 tcg acg cca agg ccc aag aaa tcc cag aac tct ctt gcg ccc gac aca 1824Ser Thr Pro Arg Pro Lys Lys Ser Gln Asn Ser Leu Ala Pro Asp Thr 595 600 605 act cca cgc gta cgc atc cgg gaa ccg tct caa ggc cca gat ggc gat 1872Thr Pro Arg Val Arg Ile Arg Glu Pro Ser Gln Gly Pro Asp Gly Asp 610 615 620 gac gat gtg gac gag ctc aat cta caa gac ggt gaa aag cct aag aag 1920Asp Asp Val Asp Glu Leu Asn Leu Gln Asp Gly Glu Lys Pro Lys Lys 625 630 635 640 aag gct cgc cgc ggt cgt cgt ggt ggc aag aat cat agg cgg ggc aag 1968Lys Ala Arg Arg Gly Arg Arg Gly Gly Lys Asn His Arg Arg Gly Lys 645 650 655 aag ccc aat agc gac agc gaa tcc agg gac ccg gcc gat cgc gtt gtt 2016Lys Pro Asn Ser Asp Ser Glu Ser Arg Asp Pro Ala Asp Arg Val Val 660 665 670 gat gaa gtg aac aag ctt caa cct cag cct cgc ttg gaa ccc gat gta 2064Asp Glu Val Asn Lys Leu Gln Pro Gln Pro Arg Leu Glu Pro Asp Val 675 680 685 cag ctg gcc cgg acg gtg tcg cat gag atc atg gaa atg gat ggc gtt 2112Gln Leu Ala Arg Thr Val Ser His Glu Ile Met Glu Met Asp Gly Val 690 695 700 ctc cag atc ggc cgt ctt agg gtg ttc act gac gtg gtc ctg gga cac 2160Leu Gln Ile Gly Arg Leu Arg Val Phe Thr Asp Val Val Leu Gly His 705 710 715 720 ggc agc cac ggg acc gtg gtg tat cgg ggc tcg ttc gat gga cgc gac 2208Gly Ser His Gly Thr Val Val Tyr Arg Gly Ser Phe Asp Gly Arg Asp 725 730 735 gtg gct gtc aag cgc atg ctg gta gaa ttc tat gat att gca tcc cat 2256Val Ala Val Lys Arg Met Leu Val Glu Phe Tyr Asp Ile Ala Ser His 740 745 750 gaa gtg ggc ctg ttg caa gaa agt gat gac cat ggc aat gtg atc cgg 2304Glu Val Gly Leu Leu Gln Glu Ser Asp Asp His Gly Asn Val Ile Arg 755 760 765 tac tac tgc cga gag cag gct gct ggt ttc ctc tac att gct ttg gag 2352Tyr Tyr Cys Arg Glu Gln Ala Ala Gly Phe Leu Tyr Ile Ala Leu Glu 770 775 780 ctc tgc ccg gcc tct ttg cag gat gtg gtt gaa cgt cca tca gat ttc 2400Leu Cys Pro Ala Ser Leu Gln Asp Val Val Glu Arg Pro Ser Asp Phe 785 790 795 800 ccg cag tta gtc cag ggc ggc ttg gac ctg ccg gac gtt ctg cgc cag 2448Pro Gln Leu Val Gln Gly Gly Leu Asp Leu Pro Asp Val Leu Arg Gln 805 810 815 att gtg gca ggt gtt cgc tat ctt cat tct ctt aag att gtg cac cgc 2496Ile Val Ala Gly Val Arg Tyr Leu His Ser Leu Lys Ile Val His Arg 820 825 830 gat ctg aag cca cag aac atc ttg gtg gcg atg cct cgc ggg cgt act 2544Asp Leu Lys Pro Gln Asn Ile Leu Val Ala Met Pro Arg Gly Arg Thr 835 840 845 ggt tca cgc tcc ctg cgg ttg ctg atc tcg gat ttc ggc ttg tgt aag

2592Gly Ser Arg Ser Leu Arg Leu Leu Ile Ser Asp Phe Gly Leu Cys Lys 850 855 860 aag ctc gaa gac aac cag agc tcc ttc cgc gca act acg gca cat gcc 2640Lys Leu Glu Asp Asn Gln Ser Ser Phe Arg Ala Thr Thr Ala His Ala 865 870 875 880 gcg ggt acc tca ggc tgg cga gcc cct gaa ttg ctg gta gac gac gac 2688Ala Gly Thr Ser Gly Trp Arg Ala Pro Glu Leu Leu Val Asp Asp Asp 885 890 895 atg agc ccg gct atg cag ggt agc gag tcc caa cac acc gaa tca tca 2736Met Ser Pro Ala Met Gln Gly Ser Glu Ser Gln His Thr Glu Ser Ser 900 905 910 gaa cca gct gtg gtg gat cct caa acc aac cgg cgg gct act cga gct 2784Glu Pro Ala Val Val Asp Pro Gln Thr Asn Arg Arg Ala Thr Arg Ala 915 920 925 atc gac atc ttc tct ttg ggc tgc gtc ttt tat tac gtt ctg acg cgg 2832Ile Asp Ile Phe Ser Leu Gly Cys Val Phe Tyr Tyr Val Leu Thr Arg 930 935 940 ggg tgc cat cct ttt gac aag aat ggc aag ttt atg cgc gag gcc aac 2880Gly Cys His Pro Phe Asp Lys Asn Gly Lys Phe Met Arg Glu Ala Asn 945 950 955 960 att gtc aag ggc aac cac aac ctc gat gag ctg cag cgt ctg ggc gac 2928Ile Val Lys Gly Asn His Asn Leu Asp Glu Leu Gln Arg Leu Gly Asp 965 970 975 tat gcc tac gag gct gaa gat cta atc cag tcc atg ttg tcg ctt gat 2976Tyr Ala Tyr Glu Ala Glu Asp Leu Ile Gln Ser Met Leu Ser Leu Asp 980 985 990 cct cga cga cga ccc gat gcg agc gct gtg ttg acg cac ccg ttc ttt 3024Pro Arg Arg Arg Pro Asp Ala Ser Ala Val Leu Thr His Pro Phe Phe 995 1000 1005 tgg cct cca tct gac cgt ctt agc ttc ctc tgc gat gtc tcg gat cac 3072Trp Pro Pro Ser Asp Arg Leu Ser Phe Leu Cys Asp Val Ser Asp His 1010 1015 1020 ttt gaa ttt gaa ccg cgg gat cct cct tcg gac gcc ctt ttg tgt ctc 3120Phe Glu Phe Glu Pro Arg Asp Pro Pro Ser Asp Ala Leu Leu Cys Leu 1025 1030 1035 1040gag tcg gtc gct cca cga gtg atg ggc ccg gac atg gat ttc ctg cga 3168Glu Ser Val Ala Pro Arg Val Met Gly Pro Asp Met Asp Phe Leu Arg 1045 1050 1055 cta ctg cca cgg gac ttt aag gat aat ctc ggc aag cag cgt aag tac 3216Leu Leu Pro Arg Asp Phe Lys Asp Asn Leu Gly Lys Gln Arg Lys Tyr 1060 1065 1070 acg gga tcg aag atg tta gat ttg ctg cga gcc ctc cgg aac aag cgc 3264Thr Gly Ser Lys Met Leu Asp Leu Leu Arg Ala Leu Arg Asn Lys Arg 1075 1080 1085 aac cat tac aac gac atg ccg gag cat ctc aag gca cac atc ggc ggg 3312Asn His Tyr Asn Asp Met Pro Glu His Leu Lys Ala His Ile Gly Gly 1090 1095 1100 ttg ccc gag ggg tat ctt aat ttt tgg act gtg cga ttc ccc agt ctt 3360Leu Pro Glu Gly Tyr Leu Asn Phe Trp Thr Val Arg Phe Pro Ser Leu 1105 1110 1115 1120ctc atg agc tgc cac tcc gtc att gtg gag ttg cgt ttg acg cgg tcc 3408Leu Met Ser Cys His Ser Val Ile Val Glu Leu Arg Leu Thr Arg Ser 1125 1130 1135 gac cgt ttc aag cgc tac ttc acg gcg act gac tag 3444Asp Arg Phe Lys Arg Tyr Phe Thr Ala Thr Asp 1140 1145 91147PRTAspergillus niger 9Met Arg Trp Arg Leu Pro Gly Ala Arg Ser Thr Leu Pro Ala Ser Val 1 5 10 15 Ala Leu Leu Leu Leu Pro Val Leu Val Ala Pro Gln Gln Trp His Glu 20 25 30 His Gln His Glu Leu Ser Ser Thr Val Ser Val Pro Leu Arg Pro Thr 35 40 45 Gly Phe Thr Ser Gly Val Asp Thr Pro Pro Ser Phe Asp Val Lys Ser 50 55 60 Asn Asp Ala Ser Ala Leu Ala Thr Leu Ala Leu Ala Gly Ser Gly Arg 65 70 75 80 Ala Val Arg Ala Pro Pro Ala Gln Ala Ser Ser Ser Thr Ala Gly Leu 85 90 95 Ala Pro Gln Leu His Ala Arg Ser Leu Gln Asp Trp Glu Val Glu Asp 100 105 110 Phe Val Leu Leu Ala Thr Val Asp Gly Ser Ile His Ala Arg Asp Arg 115 120 125 Lys Thr Gly Ala Ala Arg Trp Ala Leu Glu Val Pro Ser Ser Pro Met 130 135 140 Val Glu Ser Leu Tyr His Arg Ala Asn Arg Ser Ser Phe Asp Arg Ala 145 150 155 160 Gln Pro Glu Asp Asp Phe Ile Trp Ile Val Glu Pro Ser Gln Gly Gly 165 170 175 Ser Leu Tyr Ile Tyr Ser Ser Gly Pro Glu Ala Gly Leu Gln Lys Leu 180 185 190 Gly Leu Thr Val Lys Glu Leu Val Asp Glu Thr Pro Tyr Ser Gly Thr 195 200 205 Asp Pro Ala Val Thr Tyr Thr Ala Arg Lys Glu Thr Thr Leu Tyr Thr 210 215 220 Ile Asp Ala Arg Thr Gly Asn Ile Leu Arg Val Phe Ser Ser Arg Gly 225 230 235 240 Pro Ile Ser Ser Gly Gln Glu Cys Arg Lys Val Asp Gly Leu Asp Val 245 250 255 Asp Met Glu Glu Cys Glu Ser Pro Ser Gly Thr Leu Val Leu Gly Arg 260 265 270 Val Glu Tyr Thr Val Ala Ile Gln Asn Thr Glu Thr Gly Asp Pro Ile 275 280 285 Cys Thr Leu Lys Tyr Ser Glu Trp Thr Ala Asn Asn Arg Asp Met Asp 290 295 300 Leu Gln Ser Gln Tyr Leu Arg Thr Met Asp Gln Ser His Ile Tyr Ser 305 310 315 320 Met His Asp Gly Val Val Leu Gly Phe Asp His Ser Arg Met Asp Arg 325 330 335 Pro Arg Tyr Thr Gln Arg Phe Ser Ser Pro Val Val Arg Val Phe Asp 340 345 350 Val Ala Arg Pro Val Ser Ala Asp Ser Ser Asn Asp Pro Thr Pro Leu 355 360 365 Ile Leu Leu Ser Gln Pro Leu Gln Pro Pro Asp Pro Asp Tyr Gly Thr 370 375 380 Leu Asp Asp Arg Asp Glu Arg Val Phe Ile Asp Tyr Thr Glu Gly Gly 385 390 395 400 Gly Trp Tyr Ala Met Ser Glu Ala Thr Tyr Pro Leu Val Thr Gly Arg 405 410 415 Ala Lys Met Ala Gln Cys Tyr Glu Lys Asp Tyr Leu Arg His Gly Gln 420 425 430 Pro Leu Thr Ser Leu Thr Pro Ser Gln Gln Gln Asp Ala Leu Ala Gly 435 440 445 Val His Ser Leu Asn Gly Pro Arg Val Val Arg Arg His Ile Pro Ser 450 455 460 Ile Ser Gly Pro Ser Ser Ala Asp Met Ser Asn Asp Thr Pro Arg Glu 465 470 475 480 Leu Ile Tyr Ser Ser Ser Asp Leu Ala Leu Pro Pro Ala Leu Arg His 485 490 495 Ser Thr Ile Ile Arg Lys Gly Trp Asp Asn Ala Ile Asp Ile Phe Val 500 505 510 Thr Leu Leu Leu Leu Phe Phe Gly Thr Phe Ile Trp Phe Asn Ser His 515 520 525 His Ile Gln Glu Leu Ala Lys Gln Lys Leu Asp Leu Lys Asn Ile Met 530 535 540 Ala Ser Tyr Gly Gln Pro Pro Met Ser Thr Pro Ser Thr Pro Ile Val 545 550 555 560 Glu Ala Pro His Leu Lys Arg Glu Ala Ser Pro Asn Arg Met Ala Asn 565 570 575 Leu Thr Val Asp Met Asn Val Ser Gly Glu Gln Pro Gln Gly Gly Asp 580 585 590 Ser Thr Pro Arg Pro Lys Lys Ser Gln Asn Ser Leu Ala Pro Asp Thr 595 600 605 Thr Pro Arg Val Arg Ile Arg Glu Pro Ser Gln Gly Pro Asp Gly Asp 610 615 620 Asp Asp Val Asp Glu Leu Asn Leu Gln Asp Gly Glu Lys Pro Lys Lys 625 630 635 640 Lys Ala Arg Arg Gly Arg Arg Gly Gly Lys Asn His Arg Arg Gly Lys 645 650 655 Lys Pro Asn Ser Asp Ser Glu Ser Arg Asp Pro Ala Asp Arg Val Val 660 665 670 Asp Glu Val Asn Lys Leu Gln Pro Gln Pro Arg Leu Glu Pro Asp Val 675 680 685 Gln Leu Ala Arg Thr Val Ser His Glu Ile Met Glu Met Asp Gly Val 690 695 700 Leu Gln Ile Gly Arg Leu Arg Val Phe Thr Asp Val Val Leu Gly His 705 710 715 720 Gly Ser His Gly Thr Val Val Tyr Arg Gly Ser Phe Asp Gly Arg Asp 725 730 735 Val Ala Val Lys Arg Met Leu Val Glu Phe Tyr Asp Ile Ala Ser His 740 745 750 Glu Val Gly Leu Leu Gln Glu Ser Asp Asp His Gly Asn Val Ile Arg 755 760 765 Tyr Tyr Cys Arg Glu Gln Ala Ala Gly Phe Leu Tyr Ile Ala Leu Glu 770 775 780 Leu Cys Pro Ala Ser Leu Gln Asp Val Val Glu Arg Pro Ser Asp Phe 785 790 795 800 Pro Gln Leu Val Gln Gly Gly Leu Asp Leu Pro Asp Val Leu Arg Gln 805 810 815 Ile Val Ala Gly Val Arg Tyr Leu His Ser Leu Lys Ile Val His Arg 820 825 830 Asp Leu Lys Pro Gln Asn Ile Leu Val Ala Met Pro Arg Gly Arg Thr 835 840 845 Gly Ser Arg Ser Leu Arg Leu Leu Ile Ser Asp Phe Gly Leu Cys Lys 850 855 860 Lys Leu Glu Asp Asn Gln Ser Ser Phe Arg Ala Thr Thr Ala His Ala 865 870 875 880 Ala Gly Thr Ser Gly Trp Arg Ala Pro Glu Leu Leu Val Asp Asp Asp 885 890 895 Met Ser Pro Ala Met Gln Gly Ser Glu Ser Gln His Thr Glu Ser Ser 900 905 910 Glu Pro Ala Val Val Asp Pro Gln Thr Asn Arg Arg Ala Thr Arg Ala 915 920 925 Ile Asp Ile Phe Ser Leu Gly Cys Val Phe Tyr Tyr Val Leu Thr Arg 930 935 940 Gly Cys His Pro Phe Asp Lys Asn Gly Lys Phe Met Arg Glu Ala Asn 945 950 955 960 Ile Val Lys Gly Asn His Asn Leu Asp Glu Leu Gln Arg Leu Gly Asp 965 970 975 Tyr Ala Tyr Glu Ala Glu Asp Leu Ile Gln Ser Met Leu Ser Leu Asp 980 985 990 Pro Arg Arg Arg Pro Asp Ala Ser Ala Val Leu Thr His Pro Phe Phe 995 1000 1005 Trp Pro Pro Ser Asp Arg Leu Ser Phe Leu Cys Asp Val Ser Asp His 1010 1015 1020 Phe Glu Phe Glu Pro Arg Asp Pro Pro Ser Asp Ala Leu Leu Cys Leu 1025 1030 1035 1040Glu Ser Val Ala Pro Arg Val Met Gly Pro Asp Met Asp Phe Leu Arg 1045 1050 1055 Leu Leu Pro Arg Asp Phe Lys Asp Asn Leu Gly Lys Gln Arg Lys Tyr 1060 1065 1070 Thr Gly Ser Lys Met Leu Asp Leu Leu Arg Ala Leu Arg Asn Lys Arg 1075 1080 1085 Asn His Tyr Asn Asp Met Pro Glu His Leu Lys Ala His Ile Gly Gly 1090 1095 1100 Leu Pro Glu Gly Tyr Leu Asn Phe Trp Thr Val Arg Phe Pro Ser Leu 1105 1110 1115 1120Leu Met Ser Cys His Ser Val Ile Val Glu Leu Arg Leu Thr Arg Ser 1125 1130 1135 Asp Arg Phe Lys Arg Tyr Phe Thr Ala Thr Asp 1140 1145 102332DNAAspergillus niger 10tcccttcttc ctcttctccc tctcccttta ccttccccgt gcagctactt gactgttgag 60cttgccttcc tctccctctt tgatccaccc ttaccctctc cggaggttca attgctggtg 120cgctctgttc gcctttgttt gttttccttc ccttcctctc tcggacgttc tggttttaag 180agcgccggtc gctatccacc atgcgtttca acgctgcttt gacttctgcc ctggtctcct 240cggcttccat catgggctat gcccatgctg aggagaccga gaagaagccc gagaccacct 300ccttggccga gaagcctacc ttcaccgtga gtaattctgc agaatgatat gaggtgctgc 360ccgcgccctg ctaactgcgt gcttccagcc cacctccatc gaggctcctt tcttggagca 420gttcaccgat gactgggact cccggtggac tccctctcac gctaagaagg aggactccaa 480gtccgaggaa gactgggcct atgttggtga atggtccgtc gaggagccca ctgtcctcaa 540gggtatggag ggtgacaagg gtctcgtcgt caagaacgtc gctgcccacc acgccatctc 600ggctaagttt cccaagaaga tcgacaacaa ggacaagact ctggtcgtcc agtatgaggt 660gaagccgcag agtaagtgtg acaattgctg cgcatgctgg gtgctgggac actaacgata 720gtcacagact cccttgtctg cggtggtgcc tacctgaagc tcctccagga caacaagcag 780ctccacctcg acgagttctc gaacgcgtct ccctacgtga tcatgttcgg tcccgacaag 840tgtggtgcca ccaacaaggt aagatcttgc taaacgctgc ttagtgcagg tgactgcaac 900agggactaac agctctatct tttttaggtt cacttcatct tccgtcacaa gaaccccaag 960accggcgagt acgaggagaa gcaccttaag gcacctcccg ccgcccgtac ctccaaggtt 1020acctccgttt acaccctggt cgtcaacccc gatcagacct tccagatcct gattgatggc 1080gagtccgtca aggaaggttc cctccttgag gacttcaacc cccctgtcaa ccccgagaag 1140gagatcgacg accccaagga caagaagccc gccgactggg ttgatgaggc caagatcccc 1200gaccctgagg ctacgaagcc cgaggactgg gacgaggagg ctcccttcga gattgtcgac 1260gaggaggcta ccattcccga ggactggctc gaggacgagc ccactagcat ccctgaccct 1320gaggccgaga agcccgagga ctgggatgat gaggaggatg gcgactgggt tcctcccact 1380gttcccaacc ccaagtgcca ggatgcctcc ggatgtggtc cttggtctcc ccctatgaag 1440aagaaccctg actacaaggg caagtggtct gctcccttga ttgacaaccc ggcctacaag 1500ggaccctggg ccccccgcaa gattgccaac cccgcctact tcgaggacaa gactccctcc 1560aactttgagc ccatgggcgc tgtaagtgta cctcttaatt ctaatgctaa ggctttggat 1620gactaatgat gatgcagatt ggtttcgaga tttggaccat gcagaacgac atcctgttcg 1680acaacatcta cgttggtcac tccgccgagg atgccgagaa gctgcgccag gagaccttcg 1740atgtcaagca ccccattgag ctggctgagg aggaggccaa caagcccaag cctgaagaga 1800aggccgccga acccagcgtt agcttcaagg aagaccccgt gggccacatc aaggagaagg 1860tcgacaactt tgtccgcctc tccaagcagg accccatcaa cgccgtgaag caggttcctg 1920acgttgccgg tggtcttgcc gctgttctcg tcacaatgat ccttgtcatc gtcggagccg 1980ttggtgccag caccccggcc cctgcccccg ccaagaaggg caaggaggct gctggtgcta 2040ccaaggagaa gactggtgcg gcctccagct cctccgcaga cactggcaag ggtggtgcta 2100ccaagcgcac tacccgctct tctgccgagt aaagtggtgc agctatcggt ggaacagcac 2160gcagggaaga aagggggaga gtttaaaagg cgaaaaggtc aaacaaacaa acaaaccagg 2220gatatcctaa ctacattgtg tttttatttt atacctctgt tgcagcgttc aatcaatgtt 2280tcattctgat tccatggtga gaaggccagc tgggtatcag ctgccgccta ta 2332111689DNAAspergillus nigerCDS(1)..(1689) 11atg cgt ttc aac gct gct ttg act tct gcc ctg gtc tcc tcg gct tcc 48Met Arg Phe Asn Ala Ala Leu Thr Ser Ala Leu Val Ser Ser Ala Ser 1 5 10 15 atc atg ggc tat gcc cat gct gag gag acc gag aag aag ccc gag acc 96Ile Met Gly Tyr Ala His Ala Glu Glu Thr Glu Lys Lys Pro Glu Thr 20 25 30 acc tcc ttg gcc gag aag cct acc ttc acc ccc acc tcc atc gag gct 144Thr Ser Leu Ala Glu Lys Pro Thr Phe Thr Pro Thr Ser Ile Glu Ala 35 40 45 cct ttc ttg gag cag ttc acc gat gac tgg gac tcc cgg tgg act ccc 192Pro Phe Leu Glu Gln Phe Thr Asp Asp Trp Asp Ser Arg Trp Thr Pro 50 55 60 tct cac gct aag aag gag gac tcc aag tcc gag gaa gac tgg gcc tat 240Ser His Ala Lys Lys Glu Asp Ser Lys Ser Glu Glu Asp Trp Ala Tyr 65 70 75 80 gtt ggt gaa tgg tcc gtc gag gag ccc act gtc ctc aag ggt atg gag 288Val Gly Glu Trp Ser Val Glu Glu Pro Thr Val Leu Lys Gly Met Glu 85 90 95 ggt gac aag ggt ctc gtc gtc aag aac gtc gct gcc cac cac gcc atc 336Gly Asp Lys Gly Leu Val Val Lys Asn Val Ala Ala His His Ala Ile 100 105 110 tcg gct aag ttt ccc aag aag atc gac aac aag gac aag act ctg gtc 384Ser Ala Lys Phe Pro Lys Lys Ile Asp Asn Lys Asp Lys Thr Leu Val 115 120 125 gtc cag tat gag gtg aag ccg cag aac tcc ctt gtc tgc ggt ggt gcc 432Val Gln Tyr Glu Val Lys Pro Gln Asn Ser Leu Val Cys Gly Gly Ala 130 135 140 tac ctg aag ctc ctc cag gac aac aag cag ctc cac ctc gac gag ttc 480Tyr Leu Lys Leu Leu Gln Asp Asn Lys Gln Leu His Leu Asp Glu Phe 145 150 155 160 tcg aac gcg tct ccc tac gtg atc atg ttc ggt ccc gac aag tgt ggt 528Ser Asn Ala Ser Pro Tyr Val Ile Met Phe Gly Pro Asp Lys Cys Gly 165 170 175 gcc acc aac aag gtt cac ttc atc ttc cgt cac aag aac ccc aag acc 576Ala Thr Asn Lys Val His Phe Ile Phe Arg His Lys Asn Pro Lys Thr 180 185 190 ggc gag tac gag gag aag cac ctt aag gca cct ccc gcc gcc cgt acc 624Gly Glu Tyr Glu Glu Lys His Leu Lys

Ala Pro Pro Ala Ala Arg Thr 195 200 205 tcc aag gtt acc tcc gtt tac acc ctg gtc gtc aac ccc gat cag acc 672Ser Lys Val Thr Ser Val Tyr Thr Leu Val Val Asn Pro Asp Gln Thr 210 215 220 ttc cag atc ctg att gat ggc gag tcc gtc aag gaa ggt tcc ctc ctt 720Phe Gln Ile Leu Ile Asp Gly Glu Ser Val Lys Glu Gly Ser Leu Leu 225 230 235 240 gag gac ttc aac ccc cct gtc aac ccc gag aag gag atc gac gac ccc 768Glu Asp Phe Asn Pro Pro Val Asn Pro Glu Lys Glu Ile Asp Asp Pro 245 250 255 aag gac aag aag ccc gcc gac tgg gtt gat gag gcc aag atc ccc gac 816Lys Asp Lys Lys Pro Ala Asp Trp Val Asp Glu Ala Lys Ile Pro Asp 260 265 270 cct gag gct acg aag ccc gag gac tgg gac gag gag gct ccc ttc gag 864Pro Glu Ala Thr Lys Pro Glu Asp Trp Asp Glu Glu Ala Pro Phe Glu 275 280 285 att gtc gac gag gag gct acc att ccc gag gac tgg ctc gag gac gag 912Ile Val Asp Glu Glu Ala Thr Ile Pro Glu Asp Trp Leu Glu Asp Glu 290 295 300 ccc act agc atc cct gac cct gag gcc gag aag ccc gag gac tgg gat 960Pro Thr Ser Ile Pro Asp Pro Glu Ala Glu Lys Pro Glu Asp Trp Asp 305 310 315 320 gat gag gag gat ggc gac tgg gtt cct ccc act gtt ccc aac ccc aag 1008Asp Glu Glu Asp Gly Asp Trp Val Pro Pro Thr Val Pro Asn Pro Lys 325 330 335 tgc cag gat gcc tcc gga tgt ggt cct tgg tct ccc cct atg aag aag 1056Cys Gln Asp Ala Ser Gly Cys Gly Pro Trp Ser Pro Pro Met Lys Lys 340 345 350 aac cct gac tac aag ggc aag tgg tct gct ccc ttg att gac aac ccg 1104Asn Pro Asp Tyr Lys Gly Lys Trp Ser Ala Pro Leu Ile Asp Asn Pro 355 360 365 gcc tac aag gga ccc tgg gcc ccc cgc aag att gcc aac ccc gcc tac 1152Ala Tyr Lys Gly Pro Trp Ala Pro Arg Lys Ile Ala Asn Pro Ala Tyr 370 375 380 ttc gag gac aag act ccc tcc aac ttt gag ccc atg ggc gct att ggt 1200Phe Glu Asp Lys Thr Pro Ser Asn Phe Glu Pro Met Gly Ala Ile Gly 385 390 395 400 ttc gag att tgg acc atg cag aac gac atc ctg ttc gac aac atc tac 1248Phe Glu Ile Trp Thr Met Gln Asn Asp Ile Leu Phe Asp Asn Ile Tyr 405 410 415 gtt ggt cac tcc gcc gag gat gcc gag aag ctg cgc cag gag acc ttc 1296Val Gly His Ser Ala Glu Asp Ala Glu Lys Leu Arg Gln Glu Thr Phe 420 425 430 gat gtc aag cac ccc att gag ctg gct gag gag gag gcc aac aag ccc 1344Asp Val Lys His Pro Ile Glu Leu Ala Glu Glu Glu Ala Asn Lys Pro 435 440 445 aag cct gaa gag aag gcc gcc gaa ccc agc gtt agc ttc aag gaa gac 1392Lys Pro Glu Glu Lys Ala Ala Glu Pro Ser Val Ser Phe Lys Glu Asp 450 455 460 ccc gtg ggc cac atc aag gag aag gtc gac aac ttt gtc cgc ctc tcc 1440Pro Val Gly His Ile Lys Glu Lys Val Asp Asn Phe Val Arg Leu Ser 465 470 475 480 aag cag gac ccc atc aac gcc gtg aag cag gtt cct gac gtt gcc ggt 1488Lys Gln Asp Pro Ile Asn Ala Val Lys Gln Val Pro Asp Val Ala Gly 485 490 495 ggt ctt gcc gct gtt ctc gtc aca atg atc ctt gtc atc gtc gga gcc 1536Gly Leu Ala Ala Val Leu Val Thr Met Ile Leu Val Ile Val Gly Ala 500 505 510 gtt ggt gcc agc acc ccg gcc cct gcc ccc gcc aag aag ggc aag gag 1584Val Gly Ala Ser Thr Pro Ala Pro Ala Pro Ala Lys Lys Gly Lys Glu 515 520 525 gct gct ggt gct acc aag gag aag act ggt gcg gcc tcc agc tcc tcc 1632Ala Ala Gly Ala Thr Lys Glu Lys Thr Gly Ala Ala Ser Ser Ser Ser 530 535 540 gca gac act ggc aag ggt ggt gct acc aag cgc act acc cgc tct tct 1680Ala Asp Thr Gly Lys Gly Gly Ala Thr Lys Arg Thr Thr Arg Ser Ser 545 550 555 560 gcc gag taa 1689Ala Glu 12562PRTAspergillus niger 12Met Arg Phe Asn Ala Ala Leu Thr Ser Ala Leu Val Ser Ser Ala Ser 1 5 10 15 Ile Met Gly Tyr Ala His Ala Glu Glu Thr Glu Lys Lys Pro Glu Thr 20 25 30 Thr Ser Leu Ala Glu Lys Pro Thr Phe Thr Pro Thr Ser Ile Glu Ala 35 40 45 Pro Phe Leu Glu Gln Phe Thr Asp Asp Trp Asp Ser Arg Trp Thr Pro 50 55 60 Ser His Ala Lys Lys Glu Asp Ser Lys Ser Glu Glu Asp Trp Ala Tyr 65 70 75 80 Val Gly Glu Trp Ser Val Glu Glu Pro Thr Val Leu Lys Gly Met Glu 85 90 95 Gly Asp Lys Gly Leu Val Val Lys Asn Val Ala Ala His His Ala Ile 100 105 110 Ser Ala Lys Phe Pro Lys Lys Ile Asp Asn Lys Asp Lys Thr Leu Val 115 120 125 Val Gln Tyr Glu Val Lys Pro Gln Asn Ser Leu Val Cys Gly Gly Ala 130 135 140 Tyr Leu Lys Leu Leu Gln Asp Asn Lys Gln Leu His Leu Asp Glu Phe 145 150 155 160 Ser Asn Ala Ser Pro Tyr Val Ile Met Phe Gly Pro Asp Lys Cys Gly 165 170 175 Ala Thr Asn Lys Val His Phe Ile Phe Arg His Lys Asn Pro Lys Thr 180 185 190 Gly Glu Tyr Glu Glu Lys His Leu Lys Ala Pro Pro Ala Ala Arg Thr 195 200 205 Ser Lys Val Thr Ser Val Tyr Thr Leu Val Val Asn Pro Asp Gln Thr 210 215 220 Phe Gln Ile Leu Ile Asp Gly Glu Ser Val Lys Glu Gly Ser Leu Leu 225 230 235 240 Glu Asp Phe Asn Pro Pro Val Asn Pro Glu Lys Glu Ile Asp Asp Pro 245 250 255 Lys Asp Lys Lys Pro Ala Asp Trp Val Asp Glu Ala Lys Ile Pro Asp 260 265 270 Pro Glu Ala Thr Lys Pro Glu Asp Trp Asp Glu Glu Ala Pro Phe Glu 275 280 285 Ile Val Asp Glu Glu Ala Thr Ile Pro Glu Asp Trp Leu Glu Asp Glu 290 295 300 Pro Thr Ser Ile Pro Asp Pro Glu Ala Glu Lys Pro Glu Asp Trp Asp 305 310 315 320 Asp Glu Glu Asp Gly Asp Trp Val Pro Pro Thr Val Pro Asn Pro Lys 325 330 335 Cys Gln Asp Ala Ser Gly Cys Gly Pro Trp Ser Pro Pro Met Lys Lys 340 345 350 Asn Pro Asp Tyr Lys Gly Lys Trp Ser Ala Pro Leu Ile Asp Asn Pro 355 360 365 Ala Tyr Lys Gly Pro Trp Ala Pro Arg Lys Ile Ala Asn Pro Ala Tyr 370 375 380 Phe Glu Asp Lys Thr Pro Ser Asn Phe Glu Pro Met Gly Ala Ile Gly 385 390 395 400 Phe Glu Ile Trp Thr Met Gln Asn Asp Ile Leu Phe Asp Asn Ile Tyr 405 410 415 Val Gly His Ser Ala Glu Asp Ala Glu Lys Leu Arg Gln Glu Thr Phe 420 425 430 Asp Val Lys His Pro Ile Glu Leu Ala Glu Glu Glu Ala Asn Lys Pro 435 440 445 Lys Pro Glu Glu Lys Ala Ala Glu Pro Ser Val Ser Phe Lys Glu Asp 450 455 460 Pro Val Gly His Ile Lys Glu Lys Val Asp Asn Phe Val Arg Leu Ser 465 470 475 480 Lys Gln Asp Pro Ile Asn Ala Val Lys Gln Val Pro Asp Val Ala Gly 485 490 495 Gly Leu Ala Ala Val Leu Val Thr Met Ile Leu Val Ile Val Gly Ala 500 505 510 Val Gly Ala Ser Thr Pro Ala Pro Ala Pro Ala Lys Lys Gly Lys Glu 515 520 525 Ala Ala Gly Ala Thr Lys Glu Lys Thr Gly Ala Ala Ser Ser Ser Ser 530 535 540 Ala Asp Thr Gly Lys Gly Gly Ala Thr Lys Arg Thr Thr Arg Ser Ser 545 550 555 560 Ala Glu 132097DNAAspergillus niger 13gccgcggcgg tcgccgtacg taattcgtaa tcccgcatcg ggacacctca gcatctccgc 60gcgtctttct ccccctcgac tatcgccaat tcttccacct tccgtccatc ctctcgctca 120tttctttctc cttgggacta tctcctggtg agtgtatagg tcctaggcgc ggtcgttgag 180agaccgccca tctgcccact atgaggccgt ttaccgcact tgctgcgctg tgcggcttgt 240tcctgtccag caacagcttg gtctatgccg actcggctcc ctcgagctcc cctgtagctc 300tccctcgcga tttcaagcct ccccaagtgt ttaagaacgc caatcttgtc cgcaacacca 360atttggagaa gggataccta cgtgagaccg tcaatgttgt cgttgagaat gtggacaaga 420agccgcagtc cgactactac ttgtccttcc catccgacct ttacgacaag gtcggtgccc 480tagaagtccg tgataaatcg gctcctgaac agggacgctt cgaagtggaa gctactgagt 540tcgactcaag caggtaagtc gaccacaatg gcggccaatg gctggttggt ggagcacgaa 600gccacggttg cttgatctat gcatggtgga tgtatcccgg gcggtttgct gatctcctct 660ctttctttac acagggactt ccagtacttc gttgttcacc tccccaagcc tctcgcccct 720tcgtcgcaga tcactctggg catctcctac tccgccctca acaccctgaa gccccgtcct 780gcggccatca gccagaatga tcgccagtac ctcgcctatg ccttctctgc ctacgctccc 840tcggcttaca cgacaacgac ccagaagacc aagatcaagt tccccagcac caatgttccc 900gactacacct ccacggacct gacgtcgggc gcggatccag agcgccaggg tgccacctac 960acctacggac cctacgccga cgtcgctccc gagaccacct acccggccag tgtccggtac 1020gagttcacca agcccgtcat cactgccact cttctggagc gtgacctgga agtgtcccac 1080tggggcggca acctggcgac ggaagagcgc tactggctgc gcaacaacgg ctccaagctc 1140accgacaact tcaaccgcgt ggaatggacc atcagcagct accagcagct gccgtcctcc 1200gctatccgcg agctgaagat ccccctcaag cccggctccg tggaccccta cttcaccgac 1260gacattggca acgtttccac gagccgctac cgtcccggaa aggtcccgaa ccgtgacgcc 1320tccctggagc ttcgtccccg gttccccatc ttcggcggat ggaactacag cttccgcatt 1380ggctggaaca acgacctctc tgccttcctt cgcaaggctg tcaccggcgc tgattcctac 1440gtcctcaagg tccccttcat cgagggcccc aaggtttccg agggtattca gtatgagaag 1500gccgtcgtgc gcatcatcct ccccgagggt gcccggaacg tccgctacga gctcctcgag 1560aaggcgacta gcaatggtct ccccggtgcg aaccagatcc agactgagct caccagccac 1620aagactttca tggataccct aggacgcacg gcgctgactt tgaccgtgga ggagttgact 1680gatgaggccc gtgactcgca gatagtggta agtaactacc tccatacgcc ggatagatat 1740cgggaatagt ccagctcatt tttggatata ggtcacttac gactactctc tgtgggatgg 1800attgcgcaag cccgtgacca tcacggcggg gctgttcacc gtgtttgttg ccgcgtgggc 1860gattggaaat attgacgtga gtattaagaa gcggtagatg gaggttgtat catattgttt 1920cagttatacc agccagacag acagacagaa ttcaatagta gctgtttgta gacgactaga 1980attctgatag tgtgatttcg aatgattccc tccttgaata atatggagac agtctgatgc 2040agagtggtct ttgcaccagg tagtaagtgg gctatcggtt gtcagcgtca cctgaca 2097141512DNAAspergillus nigerCDS(1)..(1512) 14atg agg ccg ttt acc gca ctt gct gcg ctg tgc ggc ttg ttc ctg tcc 48Met Arg Pro Phe Thr Ala Leu Ala Ala Leu Cys Gly Leu Phe Leu Ser 1 5 10 15 agc aac agc ttg gtc tat gcc gac tcg gct ccc tcg agc tcc cct gta 96Ser Asn Ser Leu Val Tyr Ala Asp Ser Ala Pro Ser Ser Ser Pro Val 20 25 30 gct ctc cct cgc gat ttc aag cct ccc caa gtg ttt aag aac gcc aat 144Ala Leu Pro Arg Asp Phe Lys Pro Pro Gln Val Phe Lys Asn Ala Asn 35 40 45 ctt gtc cgc aac acc aat ttg gag aag gga tac cta cgt gag acc gtc 192Leu Val Arg Asn Thr Asn Leu Glu Lys Gly Tyr Leu Arg Glu Thr Val 50 55 60 aat gtt gtc gtt gag aat gtg gac aag aag ccg cag tcc gac tac tac 240Asn Val Val Val Glu Asn Val Asp Lys Lys Pro Gln Ser Asp Tyr Tyr 65 70 75 80 ttg tcc ttc cca tcc gac ctt tac gac aag gtc ggt gcc cta gaa gtc 288Leu Ser Phe Pro Ser Asp Leu Tyr Asp Lys Val Gly Ala Leu Glu Val 85 90 95 cgt gat aaa tcg gct cct gaa cag gga cgc ttc gaa gtg gaa gct act 336Arg Asp Lys Ser Ala Pro Glu Gln Gly Arg Phe Glu Val Glu Ala Thr 100 105 110 gag ttc gac tca agc agg gac ttc cag tac ttc gtt gtt cac ctc ccc 384Glu Phe Asp Ser Ser Arg Asp Phe Gln Tyr Phe Val Val His Leu Pro 115 120 125 aag cct ctc gcc cct tcg tcg cag atc act ctg ggc atc tcc tac tcc 432Lys Pro Leu Ala Pro Ser Ser Gln Ile Thr Leu Gly Ile Ser Tyr Ser 130 135 140 gcc ctc aac acc ctg aag ccc cgt cct gcg gcc atc agc cag aat gat 480Ala Leu Asn Thr Leu Lys Pro Arg Pro Ala Ala Ile Ser Gln Asn Asp 145 150 155 160 cgc cag tac ctc gcc tat gcc ttc tct gcc tac gct ccc tcg gct tac 528Arg Gln Tyr Leu Ala Tyr Ala Phe Ser Ala Tyr Ala Pro Ser Ala Tyr 165 170 175 acg aca acg acc cag aag acc aag atc aag ttc ccc agc acc aat gtt 576Thr Thr Thr Thr Gln Lys Thr Lys Ile Lys Phe Pro Ser Thr Asn Val 180 185 190 ccc gac tac acc tcc acg gac ctg acg tcg ggc gcg gat cca gag cgc 624Pro Asp Tyr Thr Ser Thr Asp Leu Thr Ser Gly Ala Asp Pro Glu Arg 195 200 205 cag ggt gcc acc tac acc tac gga ccc tac gcc gac gtc gct ccc gag 672Gln Gly Ala Thr Tyr Thr Tyr Gly Pro Tyr Ala Asp Val Ala Pro Glu 210 215 220 acc acc tac ccg gcc agt gtc cgg tac gag ttc acc aag ccc gtc atc 720Thr Thr Tyr Pro Ala Ser Val Arg Tyr Glu Phe Thr Lys Pro Val Ile 225 230 235 240 act gcc act ctt ctg gag cgt gac ctg gaa gtg tcc cac tgg ggc ggc 768Thr Ala Thr Leu Leu Glu Arg Asp Leu Glu Val Ser His Trp Gly Gly 245 250 255 aac ctg gcg acg gaa gag cgc tac tgg ctg cgc aac aac ggc tcc aag 816Asn Leu Ala Thr Glu Glu Arg Tyr Trp Leu Arg Asn Asn Gly Ser Lys 260 265 270 ctc acc gac aac ttc aac cgc gtg gaa tgg acc atc agc agc tac cag 864Leu Thr Asp Asn Phe Asn Arg Val Glu Trp Thr Ile Ser Ser Tyr Gln 275 280 285 cag ctg ccg tcc tcc gct atc cgc gag ctg aag atc ccc ctc aag ccc 912Gln Leu Pro Ser Ser Ala Ile Arg Glu Leu Lys Ile Pro Leu Lys Pro 290 295 300 ggc tcc gtg gac ccc tac ttc acc gac gac att ggc aac gtt tcc acg 960Gly Ser Val Asp Pro Tyr Phe Thr Asp Asp Ile Gly Asn Val Ser Thr 305 310 315 320 agc cgc tac cgt ccc gga aag gtc ccg aac cgt gac gcc tcc ctg gag 1008Ser Arg Tyr Arg Pro Gly Lys Val Pro Asn Arg Asp Ala Ser Leu Glu 325 330 335 ctt cgt ccc cgg ttc ccc atc ttc ggc gga tgg aac tac agc ttc cgc 1056Leu Arg Pro Arg Phe Pro Ile Phe Gly Gly Trp Asn Tyr Ser Phe Arg 340 345 350 att ggc tgg aac aac gac ctc tct gcc ttc ctt cgc aag gct gtc acc 1104Ile Gly Trp Asn Asn Asp Leu Ser Ala Phe Leu Arg Lys Ala Val Thr 355 360 365 ggc gct gat tcc tac gtc ctc aag gtc ccc ttc atc gag ggc ccc aag 1152Gly Ala Asp Ser Tyr Val Leu Lys Val Pro Phe Ile Glu Gly Pro Lys 370 375 380 gtt tcc gag ggt att cag tat gag aag gcc gtc gtg cgc atc atc ctc 1200Val Ser Glu Gly Ile Gln Tyr Glu Lys Ala Val Val Arg Ile Ile Leu 385 390 395 400 ccc gag ggt gcc cgg aac gtc cgc tac gag ctc ctc gag aag gcg act 1248Pro Glu Gly Ala Arg Asn Val Arg Tyr Glu Leu Leu Glu Lys Ala Thr 405 410 415 agc aat ggt ctc ccc ggt gcg aac cag atc cag act gag ctc acc agc 1296Ser Asn Gly Leu Pro Gly Ala Asn Gln Ile Gln Thr Glu Leu Thr Ser 420 425 430 cac aag act ttc atg gat acc cta gga cgc acg gcg ctg act ttg acc 1344His Lys Thr Phe Met Asp Thr Leu Gly Arg Thr Ala Leu Thr Leu Thr 435 440 445 gtg gag gag ttg act gat gag gcc cgt gac tcg cag ata gtg gtc act 1392Val Glu Glu Leu Thr Asp Glu Ala Arg Asp Ser Gln Ile Val Val Thr 450 455 460 tac gac tac tct ctg tgg gat gga ttg cgc aag ccc gtg acc atc acg 1440Tyr Asp Tyr Ser Leu Trp Asp Gly Leu Arg Lys Pro Val Thr Ile Thr 465 470 475 480 gcg ggg ctg ttc acc gtg ttt gtt gcc gcg tgg gcg att gga aat att 1488Ala Gly Leu Phe Thr Val Phe Val Ala Ala Trp Ala Ile Gly Asn Ile 485 490 495 gac gtg agt att aag aag cgg tag 1512Asp Val Ser Ile Lys Lys Arg 500 15503PRTAspergillus niger 15Met Arg Pro Phe Thr Ala Leu Ala Ala Leu Cys Gly Leu Phe Leu Ser 1 5 10 15 Ser Asn Ser Leu Val Tyr Ala Asp Ser Ala Pro Ser Ser Ser Pro Val 20

25 30 Ala Leu Pro Arg Asp Phe Lys Pro Pro Gln Val Phe Lys Asn Ala Asn 35 40 45 Leu Val Arg Asn Thr Asn Leu Glu Lys Gly Tyr Leu Arg Glu Thr Val 50 55 60 Asn Val Val Val Glu Asn Val Asp Lys Lys Pro Gln Ser Asp Tyr Tyr 65 70 75 80 Leu Ser Phe Pro Ser Asp Leu Tyr Asp Lys Val Gly Ala Leu Glu Val 85 90 95 Arg Asp Lys Ser Ala Pro Glu Gln Gly Arg Phe Glu Val Glu Ala Thr 100 105 110 Glu Phe Asp Ser Ser Arg Asp Phe Gln Tyr Phe Val Val His Leu Pro 115 120 125 Lys Pro Leu Ala Pro Ser Ser Gln Ile Thr Leu Gly Ile Ser Tyr Ser 130 135 140 Ala Leu Asn Thr Leu Lys Pro Arg Pro Ala Ala Ile Ser Gln Asn Asp 145 150 155 160 Arg Gln Tyr Leu Ala Tyr Ala Phe Ser Ala Tyr Ala Pro Ser Ala Tyr 165 170 175 Thr Thr Thr Thr Gln Lys Thr Lys Ile Lys Phe Pro Ser Thr Asn Val 180 185 190 Pro Asp Tyr Thr Ser Thr Asp Leu Thr Ser Gly Ala Asp Pro Glu Arg 195 200 205 Gln Gly Ala Thr Tyr Thr Tyr Gly Pro Tyr Ala Asp Val Ala Pro Glu 210 215 220 Thr Thr Tyr Pro Ala Ser Val Arg Tyr Glu Phe Thr Lys Pro Val Ile 225 230 235 240 Thr Ala Thr Leu Leu Glu Arg Asp Leu Glu Val Ser His Trp Gly Gly 245 250 255 Asn Leu Ala Thr Glu Glu Arg Tyr Trp Leu Arg Asn Asn Gly Ser Lys 260 265 270 Leu Thr Asp Asn Phe Asn Arg Val Glu Trp Thr Ile Ser Ser Tyr Gln 275 280 285 Gln Leu Pro Ser Ser Ala Ile Arg Glu Leu Lys Ile Pro Leu Lys Pro 290 295 300 Gly Ser Val Asp Pro Tyr Phe Thr Asp Asp Ile Gly Asn Val Ser Thr 305 310 315 320 Ser Arg Tyr Arg Pro Gly Lys Val Pro Asn Arg Asp Ala Ser Leu Glu 325 330 335 Leu Arg Pro Arg Phe Pro Ile Phe Gly Gly Trp Asn Tyr Ser Phe Arg 340 345 350 Ile Gly Trp Asn Asn Asp Leu Ser Ala Phe Leu Arg Lys Ala Val Thr 355 360 365 Gly Ala Asp Ser Tyr Val Leu Lys Val Pro Phe Ile Glu Gly Pro Lys 370 375 380 Val Ser Glu Gly Ile Gln Tyr Glu Lys Ala Val Val Arg Ile Ile Leu 385 390 395 400 Pro Glu Gly Ala Arg Asn Val Arg Tyr Glu Leu Leu Glu Lys Ala Thr 405 410 415 Ser Asn Gly Leu Pro Gly Ala Asn Gln Ile Gln Thr Glu Leu Thr Ser 420 425 430 His Lys Thr Phe Met Asp Thr Leu Gly Arg Thr Ala Leu Thr Leu Thr 435 440 445 Val Glu Glu Leu Thr Asp Glu Ala Arg Asp Ser Gln Ile Val Val Thr 450 455 460 Tyr Asp Tyr Ser Leu Trp Asp Gly Leu Arg Lys Pro Val Thr Ile Thr 465 470 475 480 Ala Gly Leu Phe Thr Val Phe Val Ala Ala Trp Ala Ile Gly Asn Ile 485 490 495 Asp Val Ser Ile Lys Lys Arg 500 162212DNAAspergillus niger 16cttttctctt tcggtagctt ctgctctacg cgtcacctgc cttcctctct ctccccaccc 60ctctccctcc aaacgggccg gtctattatt tgttttcatt gactttcgag gatctccccg 120gaagccactg gaaaagcaga tatccattta aataccctct cccatcgtcc tctattccgc 180tgctgcttct ttattacaag atgcgctcct tcgcgccttg gctcgttagc cttctcggag 240catccgcggt ggttgcggct gctgataccg agtctgatgt tatttcactg gatcaggaca 300catttgagag cttcatgaac gagcacggtc tcgtgcttgc cgaattcttt gctccttggt 360gtggccactg taaagccctc gcaccaaagt atgaggaagc agctacggag ctcaaggcga 420agaatatccc tctggtgaag gttgactgca ccgccgagga ggatctctgc cggagtcagg 480gcgttgaagg ctaccctacc ctcaagatct ttcggggtgt tgactctagc aagccttacc 540agggcgctag gcaaacagaa tcgtgagtcc tcttggtttt gactggacga aagaaatgga 600tatgcatcaa aaggctatcg tgtatgtgat gtgctactga cccatcggac ggtttcccag 660aatcgtttcc tacatgatta agcagtcact tcctgcagta tcctccgtga acgaggagaa 720tttagaagag atcaagacca tggacaagat tgtcgtgatc ggttatatcc cgtccgagga 780ccaggaaact tatcaagcat tcgaaaaata tgctgagtct cagcgggata actacctctt 840tgctgccacg gacgatgccg ccattgcgaa atcggaaggt gtcgagcagc cctccatcgt 900gctctataag gacttcgatg agaagaaggc tgtttacgat ggcgagatcg aacaggaggc 960tattcacagc tgggtgaaat ccgctagtac tccccttgtg ggcgagattg gccctgagac 1020ctactctggc tatattgggg taagttgaat ctatacgtcg acgtgacatc tctttgcatt 1080tcggcttgag tttcgatacc aatttgctgg ctgaactggc taaccacttt tcttataggc 1140tggagtccca ttggcctata tctttgccga gaccaaggag gagcgcgaaa agtacaccga 1200agacttcaag cctattgccc agaagcacaa gggtgctatc aacattgcta ctattgacgc 1260caagatgttc ggtgcccacg ctggaaacct caacctagac tctcagaagt tcccggcatt 1320cgccatccag gatcccgcaa agaacgccaa atacccctat gaccaggcca aggaattgaa 1380tgccgacgag gttgaaaagt tcatccagga tgttctggat gggaaggtcg agcctagcat 1440caagtcggaa cctgttcccg aatctcagga gggccccgtc acggttgtag tggcccattc 1500ctacaaggat ctcgtcattg acaatgacaa ggatgtcttg ctcgaattct acgcaccttg 1560gtgtggacac tgcaaagcgt atgtctcttc gatcccctaa gtacactagg tgttggagca 1620caatcaacta acagcaaatc tatagtcttg ctccgaagta cgatgagctc gcagctctct 1680atgctgacca ccccgatttg gcggctaagg tcaccatcgc taagatcgat gcgacggcca 1740acgatgttcc ggacccgatt accggattcc ctaccctcag actctacccg gccggtgcca 1800aggactcccc cattgagtac tctggctcgc gcactgtcga ggatcttgcc aactttgtga 1860aggagaatgg caaacacaac gttgacgccc tcaatgtcgc ttccgaggaa acacaggagg 1920gtggtgatgt gactgaggct gctccctccg ctacggaggc cgagaccccg gctgccacag 1980atgacgagaa ggcagaacat gacgaactgt aaacagtctc ccaattgaga tcccgcttag 2040tgctgtgccg tatcaatcac taattatttg gaacttcgtt tctctattgt taacttttgt 2100aatctctgga ccaatctgtt gttggttgaa ttagctaaat tggaccagtt tctatatggc 2160cagacaatgt gatgaatata taatttgctc cacgcatgtc tacctcgttc ca 2212171548DNAAspergillus nigerCDS(1)..(1548) 17atg cgc tcc ttc gcg cct tgg ctc gtt agc ctt ctc gga gca tcc gcg 48Met Arg Ser Phe Ala Pro Trp Leu Val Ser Leu Leu Gly Ala Ser Ala 1 5 10 15 gtg gtt gcg gct gct gat acc gag tct gat gtt att tca ctg gat cag 96Val Val Ala Ala Ala Asp Thr Glu Ser Asp Val Ile Ser Leu Asp Gln 20 25 30 gac aca ttt gag agc ttc atg aac gag cac ggt ctc gtg ctt gcc gaa 144Asp Thr Phe Glu Ser Phe Met Asn Glu His Gly Leu Val Leu Ala Glu 35 40 45 ttc ttt gct cct tgg tgt ggc cac tgt aaa gcc ctc gca cca aag tat 192Phe Phe Ala Pro Trp Cys Gly His Cys Lys Ala Leu Ala Pro Lys Tyr 50 55 60 gag gaa gca gct acg gag ctc aag gcg aag aat atc cct ctg gtg aag 240Glu Glu Ala Ala Thr Glu Leu Lys Ala Lys Asn Ile Pro Leu Val Lys 65 70 75 80 gtt gac tgc acc gcc gag gag gat ctc tgc cgg agt cag ggc gtt gaa 288Val Asp Cys Thr Ala Glu Glu Asp Leu Cys Arg Ser Gln Gly Val Glu 85 90 95 ggc tac cct acc ctc aag atc ttt cgg ggt gtt gac tct agc aag cct 336Gly Tyr Pro Thr Leu Lys Ile Phe Arg Gly Val Asp Ser Ser Lys Pro 100 105 110 tac cag ggc gct agg caa aca gaa tca atc gtt tcc tac atg att aag 384Tyr Gln Gly Ala Arg Gln Thr Glu Ser Ile Val Ser Tyr Met Ile Lys 115 120 125 cag tca ctt cct gca gta tcc tcc gtg aac gag gag aat tta gaa gag 432Gln Ser Leu Pro Ala Val Ser Ser Val Asn Glu Glu Asn Leu Glu Glu 130 135 140 atc aag acc atg gac aag att gtc gtg atc ggt tat atc ccg tcc gag 480Ile Lys Thr Met Asp Lys Ile Val Val Ile Gly Tyr Ile Pro Ser Glu 145 150 155 160 gac cag gaa act tat caa gca ttc gaa aaa tat gct gag tct cag cgg 528Asp Gln Glu Thr Tyr Gln Ala Phe Glu Lys Tyr Ala Glu Ser Gln Arg 165 170 175 gat aac tac ctc ttt gct gcc acg gac gat gcc gcc att gcg aaa tcg 576Asp Asn Tyr Leu Phe Ala Ala Thr Asp Asp Ala Ala Ile Ala Lys Ser 180 185 190 gaa ggt gtc gag cag ccc tcc atc gtg ctc tat aag gac ttc gat gag 624Glu Gly Val Glu Gln Pro Ser Ile Val Leu Tyr Lys Asp Phe Asp Glu 195 200 205 aag aag gct gtt tac gat ggc gag atc gaa cag gag gct att cac agc 672Lys Lys Ala Val Tyr Asp Gly Glu Ile Glu Gln Glu Ala Ile His Ser 210 215 220 tgg gtg aaa tcc gct agt act ccc ctt gtg ggc gag att ggc cct gag 720Trp Val Lys Ser Ala Ser Thr Pro Leu Val Gly Glu Ile Gly Pro Glu 225 230 235 240 acc tac tct ggc tat att ggg gct gga gtc cca ttg gcc tat atc ttt 768Thr Tyr Ser Gly Tyr Ile Gly Ala Gly Val Pro Leu Ala Tyr Ile Phe 245 250 255 gcc gag acc aag gag gag cgc gaa aag tac acc gaa gac ttc aag cct 816Ala Glu Thr Lys Glu Glu Arg Glu Lys Tyr Thr Glu Asp Phe Lys Pro 260 265 270 att gcc cag aag cac aag ggt gct atc aac att gct act att gac gcc 864Ile Ala Gln Lys His Lys Gly Ala Ile Asn Ile Ala Thr Ile Asp Ala 275 280 285 aag atg ttc ggt gcc cac gct gga aac ctc aac cta gac tct cag aag 912Lys Met Phe Gly Ala His Ala Gly Asn Leu Asn Leu Asp Ser Gln Lys 290 295 300 ttc ccg gca ttc gcc atc cag gat ccc gca aag aac gcc aaa tac ccc 960Phe Pro Ala Phe Ala Ile Gln Asp Pro Ala Lys Asn Ala Lys Tyr Pro 305 310 315 320 tat gac cag gcc aag gaa ttg aat gcc gac gag gtt gaa aag ttc atc 1008Tyr Asp Gln Ala Lys Glu Leu Asn Ala Asp Glu Val Glu Lys Phe Ile 325 330 335 cag gat gtt ctg gat ggg aag gtc gag cct agc atc aag tcg gaa cct 1056Gln Asp Val Leu Asp Gly Lys Val Glu Pro Ser Ile Lys Ser Glu Pro 340 345 350 gtt ccc gaa tct cag gag ggc ccc gtc acg gtt gta gtg gcc cat tcc 1104Val Pro Glu Ser Gln Glu Gly Pro Val Thr Val Val Val Ala His Ser 355 360 365 tac aag gat ctc gtc att gac aat gac aag gat gtc ttg ctc gaa ttc 1152Tyr Lys Asp Leu Val Ile Asp Asn Asp Lys Asp Val Leu Leu Glu Phe 370 375 380 tac gca cct tgg tgt gga cac tgc aaa gct ctt gct ccg aag tac gat 1200Tyr Ala Pro Trp Cys Gly His Cys Lys Ala Leu Ala Pro Lys Tyr Asp 385 390 395 400 gag ctc gca gct ctc tat gct gac cac ccc gat ttg gcg gct aag gtc 1248Glu Leu Ala Ala Leu Tyr Ala Asp His Pro Asp Leu Ala Ala Lys Val 405 410 415 acc atc gct aag atc gat gcg acg gcc aac gat gtt ccg gac ccg att 1296Thr Ile Ala Lys Ile Asp Ala Thr Ala Asn Asp Val Pro Asp Pro Ile 420 425 430 acc gga ttc cct acc ctc aga ctc tac ccg gcc ggt gcc aag gac tcc 1344Thr Gly Phe Pro Thr Leu Arg Leu Tyr Pro Ala Gly Ala Lys Asp Ser 435 440 445 ccc att gag tac tct ggc tcg cgc act gtc gag gat ctt gcc aac ttt 1392Pro Ile Glu Tyr Ser Gly Ser Arg Thr Val Glu Asp Leu Ala Asn Phe 450 455 460 gtg aag gag aat ggc aaa cac aac gtt gac gcc ctc aat gtc gct tcc 1440Val Lys Glu Asn Gly Lys His Asn Val Asp Ala Leu Asn Val Ala Ser 465 470 475 480 gag gaa aca cag gag ggt ggt gat gtg act gag gct gct ccc tcc gct 1488Glu Glu Thr Gln Glu Gly Gly Asp Val Thr Glu Ala Ala Pro Ser Ala 485 490 495 acg gag gcc gag acc ccg gct gcc aca gat gac gag aag gca gaa cat 1536Thr Glu Ala Glu Thr Pro Ala Ala Thr Asp Asp Glu Lys Ala Glu His 500 505 510 gac gaa ctg taa 1548Asp Glu Leu 515 18515PRTAspergillus niger 18Met Arg Ser Phe Ala Pro Trp Leu Val Ser Leu Leu Gly Ala Ser Ala 1 5 10 15 Val Val Ala Ala Ala Asp Thr Glu Ser Asp Val Ile Ser Leu Asp Gln 20 25 30 Asp Thr Phe Glu Ser Phe Met Asn Glu His Gly Leu Val Leu Ala Glu 35 40 45 Phe Phe Ala Pro Trp Cys Gly His Cys Lys Ala Leu Ala Pro Lys Tyr 50 55 60 Glu Glu Ala Ala Thr Glu Leu Lys Ala Lys Asn Ile Pro Leu Val Lys 65 70 75 80 Val Asp Cys Thr Ala Glu Glu Asp Leu Cys Arg Ser Gln Gly Val Glu 85 90 95 Gly Tyr Pro Thr Leu Lys Ile Phe Arg Gly Val Asp Ser Ser Lys Pro 100 105 110 Tyr Gln Gly Ala Arg Gln Thr Glu Ser Ile Val Ser Tyr Met Ile Lys 115 120 125 Gln Ser Leu Pro Ala Val Ser Ser Val Asn Glu Glu Asn Leu Glu Glu 130 135 140 Ile Lys Thr Met Asp Lys Ile Val Val Ile Gly Tyr Ile Pro Ser Glu 145 150 155 160 Asp Gln Glu Thr Tyr Gln Ala Phe Glu Lys Tyr Ala Glu Ser Gln Arg 165 170 175 Asp Asn Tyr Leu Phe Ala Ala Thr Asp Asp Ala Ala Ile Ala Lys Ser 180 185 190 Glu Gly Val Glu Gln Pro Ser Ile Val Leu Tyr Lys Asp Phe Asp Glu 195 200 205 Lys Lys Ala Val Tyr Asp Gly Glu Ile Glu Gln Glu Ala Ile His Ser 210 215 220 Trp Val Lys Ser Ala Ser Thr Pro Leu Val Gly Glu Ile Gly Pro Glu 225 230 235 240 Thr Tyr Ser Gly Tyr Ile Gly Ala Gly Val Pro Leu Ala Tyr Ile Phe 245 250 255 Ala Glu Thr Lys Glu Glu Arg Glu Lys Tyr Thr Glu Asp Phe Lys Pro 260 265 270 Ile Ala Gln Lys His Lys Gly Ala Ile Asn Ile Ala Thr Ile Asp Ala 275 280 285 Lys Met Phe Gly Ala His Ala Gly Asn Leu Asn Leu Asp Ser Gln Lys 290 295 300 Phe Pro Ala Phe Ala Ile Gln Asp Pro Ala Lys Asn Ala Lys Tyr Pro 305 310 315 320 Tyr Asp Gln Ala Lys Glu Leu Asn Ala Asp Glu Val Glu Lys Phe Ile 325 330 335 Gln Asp Val Leu Asp Gly Lys Val Glu Pro Ser Ile Lys Ser Glu Pro 340 345 350 Val Pro Glu Ser Gln Glu Gly Pro Val Thr Val Val Val Ala His Ser 355 360 365 Tyr Lys Asp Leu Val Ile Asp Asn Asp Lys Asp Val Leu Leu Glu Phe 370 375 380 Tyr Ala Pro Trp Cys Gly His Cys Lys Ala Leu Ala Pro Lys Tyr Asp 385 390 395 400 Glu Leu Ala Ala Leu Tyr Ala Asp His Pro Asp Leu Ala Ala Lys Val 405 410 415 Thr Ile Ala Lys Ile Asp Ala Thr Ala Asn Asp Val Pro Asp Pro Ile 420 425 430 Thr Gly Phe Pro Thr Leu Arg Leu Tyr Pro Ala Gly Ala Lys Asp Ser 435 440 445 Pro Ile Glu Tyr Ser Gly Ser Arg Thr Val Glu Asp Leu Ala Asn Phe 450 455 460 Val Lys Glu Asn Gly Lys His Asn Val Asp Ala Leu Asn Val Ala Ser 465 470 475 480 Glu Glu Thr Gln Glu Gly Gly Asp Val Thr Glu Ala Ala Pro Ser Ala 485 490 495 Thr Glu Ala Glu Thr Pro Ala Ala Thr Asp Asp Glu Lys Ala Glu His 500 505 510 Asp Glu Leu 515 192113DNAAspergillus niger 19cccgcaatcc ccgtcgacct catcgcttcc tccctttctc ctccatcctc tctctcttcc 60gtcgtctttt cttcttctcc ttctcctttt gtacttcccc tccattcctt cagctggttc 120tcgcctccag ctttcctttc tttctttccc tcccctttta ttcgagtaat cctgcagctc 180tgggaggtgc aacagtcaca atgagcggac gtgagtcttg cacgcgatcg ctgccatctc 240cgcgacagcg ttccatcctt tacctcaatg gatcagcaaa tgctgatact cgattctagt 300ccggtttctc gatctcatca agcccttcac gcccctcctc ccggaggtgg ccgccccgga 360aaccaaggtt cccttcaacc agaagttgat gtggacgggg gtacgtgata cttgtccagc 420tcgacatgag cttctaagct aatggattac ccctgcagtt gaccctattg atcttcctgg 480tcatgagcca gatgcccttg tacggaattg tctcctctga cacctccgac cctctgtact 540ggctccgtat gatgttggcc agtaaccggg gtaccctgat ggaactgggt atcaccccca 600tcatctcctc tggcatggtt ttccaggtat gtaatgggga aattgcaatc tgatcacgga 660tatcgggcat ttgctaatat gtggcttttg tctgatagct tctcgctggt acccacctca 720tcgatgtcaa cctggacctg aagaccgacc gtgaactgta tcagaccgct cagaagctct 780tcgctatcat cctgtccttc ggtcaggcct gcgtctacgt cctcactggt ctttacggcc 840agcccagtga ccttggtgcc ggtatctgtg ttctgctgat tgttcagctg gtcgttgctg 900gcttggttgt

catcctgctg gatgagctgc tccagaaggg ctatggtctt ggtagcggta 960tctctctgtt catcgcgacc aacatctgcg agtcgatcgt ctggaaggct ttctctccta 1020cgaccatcaa cactggccgt ggtcccgagt ttgagggtgc catcattgcc ctcttccacc 1080ttctgttgac ctggtccgac aagcagcgcg ctctccgcga ggctttctac cgccagaacc 1140tccccaacat catgaacctg ctggctactc tcctcgtttt cgccgctgtg atctacctcc 1200agggcttccg tgttgagatc cctgtcaagt cctcccgcca gcgtggcatg cgtggttcct 1260accctgttcg cctgttctac acctccaaca tgcccatcat gcttcagtct gctctgtgct 1320ccaacatctt cctcatcagt cagatgctgt actctcgctt ctctgacaac ctccttgtca 1380agcttctcgg tgtttgggag cctcgtgagg gttctgccca gctccacgcc gcctccggca 1440ttgcctacta catgtctcct cccctgaact tcaaggaggc ccttcttgac cccattcaca 1500ccgccgttta catcaccttc atgctggttg cttgtgctct cttctccaag acctggattg 1560aggtttccgg ctctgctccc cgcgatgttg ccaagcagct caaggaccag ggtctcgtga 1620tggctggtca ccgtgagcag agcatgtaca aggagctcaa gcgcgtcatc cctactgctg 1680ctgctttcgg tggtgcctgc attggtgccc tgtccgtcgc ttctgacctg cttggtgctc 1740ttggcagcgg tactggtatc ctccttgccg ttacgtaagt cttcactttg gtctcagatt 1800ttctgaagtg gatactaaca ttcaaatgca ggattatata cggatacttt gaaattgccg 1860cccgtgaggg cgacattgga tcgggcctca agggccttgt tccgggtaac tagataaggc 1920cccctttttg atgaaagcat gagaagaagt ttgagggctt atgtttgttc ttgcaacttt 1980ctgtttcttc tcaggtagtg tgctgttgtg gctgggatct ggattattta gtttcttgat 2040ggatgtatgg ctagttttaa caatttgcag gaggggaaga tcttctctac ggagatacgt 2100ccacgccaca gct 2113201437DNAAspergillus nigerCDS(1)..(1437) 20atg agc gga ctc cgg ttt ctc gat ctc atc aag ccc ttc acg ccc ctc 48Met Ser Gly Leu Arg Phe Leu Asp Leu Ile Lys Pro Phe Thr Pro Leu 1 5 10 15 ctc ccg gag gtg gcc gcc ccg gaa acc aag gtt ccc ttc aac cag aag 96Leu Pro Glu Val Ala Ala Pro Glu Thr Lys Val Pro Phe Asn Gln Lys 20 25 30 ttg atg tgg acg ggg ttg acc cta ttg atc ttc ctg gtc atg agc cag 144Leu Met Trp Thr Gly Leu Thr Leu Leu Ile Phe Leu Val Met Ser Gln 35 40 45 atg ccc ttg tac gga att gtc tcc tct gac acc tcc gac cct ctg tac 192Met Pro Leu Tyr Gly Ile Val Ser Ser Asp Thr Ser Asp Pro Leu Tyr 50 55 60 tgg ctc cgt atg atg ttg gcc agt aac cgg ggt acc ctg atg gaa ctg 240Trp Leu Arg Met Met Leu Ala Ser Asn Arg Gly Thr Leu Met Glu Leu 65 70 75 80 ggt atc acc ccc atc atc tcc tct ggc atg gtt ttc cag ctt ctc gct 288Gly Ile Thr Pro Ile Ile Ser Ser Gly Met Val Phe Gln Leu Leu Ala 85 90 95 ggt acc cac ctc atc gat gtc aac ctg gac ctg aag acc gac cgt gaa 336Gly Thr His Leu Ile Asp Val Asn Leu Asp Leu Lys Thr Asp Arg Glu 100 105 110 ctg tat cag acc gct cag aag ctc ttc gct atc atc ctg tcc ttc ggt 384Leu Tyr Gln Thr Ala Gln Lys Leu Phe Ala Ile Ile Leu Ser Phe Gly 115 120 125 cag gcc tgc gtc tac gtc ctc act ggt ctt tac ggc cag ccc agt gac 432Gln Ala Cys Val Tyr Val Leu Thr Gly Leu Tyr Gly Gln Pro Ser Asp 130 135 140 ctt ggt gcc ggt atc tgt gtt ctg ctg att gtt cag ctg gtc gtt gct 480Leu Gly Ala Gly Ile Cys Val Leu Leu Ile Val Gln Leu Val Val Ala 145 150 155 160 ggc ttg gtt gtc atc ctg ctg gat gag ctg ctc cag aag ggc tat ggt 528Gly Leu Val Val Ile Leu Leu Asp Glu Leu Leu Gln Lys Gly Tyr Gly 165 170 175 ctt ggt agc ggt atc tct ctg ttc atc gcg acc aac atc tgc gag tcg 576Leu Gly Ser Gly Ile Ser Leu Phe Ile Ala Thr Asn Ile Cys Glu Ser 180 185 190 atc gtc tgg aag gct ttc tct cct acg acc atc aac act ggc cgt ggt 624Ile Val Trp Lys Ala Phe Ser Pro Thr Thr Ile Asn Thr Gly Arg Gly 195 200 205 ccc gag ttt gag ggt gcc atc att gcc ctc ttc cac ctt ctg ttg acc 672Pro Glu Phe Glu Gly Ala Ile Ile Ala Leu Phe His Leu Leu Leu Thr 210 215 220 tgg tcc gac aag cag cgc gct ctc cgc gag gct ttc tac cgc cag aac 720Trp Ser Asp Lys Gln Arg Ala Leu Arg Glu Ala Phe Tyr Arg Gln Asn 225 230 235 240 ctc ccc aac atc atg aac ctg ctg gct act ctc ctc gtt ttc gcc gct 768Leu Pro Asn Ile Met Asn Leu Leu Ala Thr Leu Leu Val Phe Ala Ala 245 250 255 gtg atc tac ctc cag ggc ttc cgt gtt gag atc cct gtc aag tcc tcc 816Val Ile Tyr Leu Gln Gly Phe Arg Val Glu Ile Pro Val Lys Ser Ser 260 265 270 cgc cag cgt ggc atg cgt ggt tcc tac cct gtt cgc ctg ttc tac acc 864Arg Gln Arg Gly Met Arg Gly Ser Tyr Pro Val Arg Leu Phe Tyr Thr 275 280 285 tcc aac atg ccc atc atg ctt cag tct gct ctg tgc tcc aac atc ttc 912Ser Asn Met Pro Ile Met Leu Gln Ser Ala Leu Cys Ser Asn Ile Phe 290 295 300 ctc atc agt cag atg ctg tac tct cgc ttc tct gac aac ctc ctt gtc 960Leu Ile Ser Gln Met Leu Tyr Ser Arg Phe Ser Asp Asn Leu Leu Val 305 310 315 320 aag ctt ctc ggt gtt tgg gag cct cgt gag ggt tct gcc cag ctc cac 1008Lys Leu Leu Gly Val Trp Glu Pro Arg Glu Gly Ser Ala Gln Leu His 325 330 335 gcc gcc tcc ggc att gcc tac tac atg tct cct ccc ctg aac ttc aag 1056Ala Ala Ser Gly Ile Ala Tyr Tyr Met Ser Pro Pro Leu Asn Phe Lys 340 345 350 gag gcc ctt ctt gac ccc att cac acc gcc gtt tac atc acc ttc atg 1104Glu Ala Leu Leu Asp Pro Ile His Thr Ala Val Tyr Ile Thr Phe Met 355 360 365 ctg gtt gct tgt gct ctc ttc tcc aag acc tgg att gag gtt tcc ggc 1152Leu Val Ala Cys Ala Leu Phe Ser Lys Thr Trp Ile Glu Val Ser Gly 370 375 380 tct gct ccc cgc gat gtt gcc aag cag ctc aag gac cag ggt ctc gtg 1200Ser Ala Pro Arg Asp Val Ala Lys Gln Leu Lys Asp Gln Gly Leu Val 385 390 395 400 atg gct ggt cac cgt gag cag agc atg tac aag gag ctc aag cgc gtc 1248Met Ala Gly His Arg Glu Gln Ser Met Tyr Lys Glu Leu Lys Arg Val 405 410 415 atc cct act gct gct gct ttc ggt ggt gcc tgc att ggt gcc ctg tcc 1296Ile Pro Thr Ala Ala Ala Phe Gly Gly Ala Cys Ile Gly Ala Leu Ser 420 425 430 gtc gct tct gac ctg ctt ggt gct ctt ggc agc ggt act ggt atc ctc 1344Val Ala Ser Asp Leu Leu Gly Ala Leu Gly Ser Gly Thr Gly Ile Leu 435 440 445 ctt gcc gtt acg att ata tac gga tac ttt gaa att gcc gcc cgt gag 1392Leu Ala Val Thr Ile Ile Tyr Gly Tyr Phe Glu Ile Ala Ala Arg Glu 450 455 460 ggc gac att gga tcg ggc ctc aag ggc ctt gtt ccg ggt aac tag 1437Gly Asp Ile Gly Ser Gly Leu Lys Gly Leu Val Pro Gly Asn 465 470 475 21478PRTAspergillus niger 21Met Ser Gly Leu Arg Phe Leu Asp Leu Ile Lys Pro Phe Thr Pro Leu 1 5 10 15 Leu Pro Glu Val Ala Ala Pro Glu Thr Lys Val Pro Phe Asn Gln Lys 20 25 30 Leu Met Trp Thr Gly Leu Thr Leu Leu Ile Phe Leu Val Met Ser Gln 35 40 45 Met Pro Leu Tyr Gly Ile Val Ser Ser Asp Thr Ser Asp Pro Leu Tyr 50 55 60 Trp Leu Arg Met Met Leu Ala Ser Asn Arg Gly Thr Leu Met Glu Leu 65 70 75 80 Gly Ile Thr Pro Ile Ile Ser Ser Gly Met Val Phe Gln Leu Leu Ala 85 90 95 Gly Thr His Leu Ile Asp Val Asn Leu Asp Leu Lys Thr Asp Arg Glu 100 105 110 Leu Tyr Gln Thr Ala Gln Lys Leu Phe Ala Ile Ile Leu Ser Phe Gly 115 120 125 Gln Ala Cys Val Tyr Val Leu Thr Gly Leu Tyr Gly Gln Pro Ser Asp 130 135 140 Leu Gly Ala Gly Ile Cys Val Leu Leu Ile Val Gln Leu Val Val Ala 145 150 155 160 Gly Leu Val Val Ile Leu Leu Asp Glu Leu Leu Gln Lys Gly Tyr Gly 165 170 175 Leu Gly Ser Gly Ile Ser Leu Phe Ile Ala Thr Asn Ile Cys Glu Ser 180 185 190 Ile Val Trp Lys Ala Phe Ser Pro Thr Thr Ile Asn Thr Gly Arg Gly 195 200 205 Pro Glu Phe Glu Gly Ala Ile Ile Ala Leu Phe His Leu Leu Leu Thr 210 215 220 Trp Ser Asp Lys Gln Arg Ala Leu Arg Glu Ala Phe Tyr Arg Gln Asn 225 230 235 240 Leu Pro Asn Ile Met Asn Leu Leu Ala Thr Leu Leu Val Phe Ala Ala 245 250 255 Val Ile Tyr Leu Gln Gly Phe Arg Val Glu Ile Pro Val Lys Ser Ser 260 265 270 Arg Gln Arg Gly Met Arg Gly Ser Tyr Pro Val Arg Leu Phe Tyr Thr 275 280 285 Ser Asn Met Pro Ile Met Leu Gln Ser Ala Leu Cys Ser Asn Ile Phe 290 295 300 Leu Ile Ser Gln Met Leu Tyr Ser Arg Phe Ser Asp Asn Leu Leu Val 305 310 315 320 Lys Leu Leu Gly Val Trp Glu Pro Arg Glu Gly Ser Ala Gln Leu His 325 330 335 Ala Ala Ser Gly Ile Ala Tyr Tyr Met Ser Pro Pro Leu Asn Phe Lys 340 345 350 Glu Ala Leu Leu Asp Pro Ile His Thr Ala Val Tyr Ile Thr Phe Met 355 360 365 Leu Val Ala Cys Ala Leu Phe Ser Lys Thr Trp Ile Glu Val Ser Gly 370 375 380 Ser Ala Pro Arg Asp Val Ala Lys Gln Leu Lys Asp Gln Gly Leu Val 385 390 395 400 Met Ala Gly His Arg Glu Gln Ser Met Tyr Lys Glu Leu Lys Arg Val 405 410 415 Ile Pro Thr Ala Ala Ala Phe Gly Gly Ala Cys Ile Gly Ala Leu Ser 420 425 430 Val Ala Ser Asp Leu Leu Gly Ala Leu Gly Ser Gly Thr Gly Ile Leu 435 440 445 Leu Ala Val Thr Ile Ile Tyr Gly Tyr Phe Glu Ile Ala Ala Arg Glu 450 455 460 Gly Asp Ile Gly Ser Gly Leu Lys Gly Leu Val Pro Gly Asn 465 470 475 225117DNAAspergillus niger 22agctcgactg ggcatcttta atgcctcatt ggtctataca ttaattatac tgaattggta 60attacattgt tgccattgat cacctcgagc ttatccgccg cccgttatgt gtgcttgcta 120gctagcttat cgggaacagc tagctagtca acaccgctgc catcagctct aggtatcatt 180tcccgtggtg ggctgtgacg atggtatctg gactggcaaa tttcgcctca tggcgacttg 240catctgtatt gattgccggc ctgctggcta tccagggacg cgctagtcca tcagtcaatg 300ttgctctcca agcttcgttt gattccccac cttatctgat agagctactg taagtgaaat 360tcagacagat ctagttggat gctcttgatt cctgggctga cctctatgta gcgaatccgc 420tgcggaggag aactccacct catacttccc gttactcgat cggatcgccg acggtatttt 480cgatgacgct gttacggata aggacctata tgatcgcttc ctggaggttg tgcgtgagga 540tggacactta cggacccctg aaagtctctc atctttcaag ctgtcgctgg cgatgagatc 600cgccagtccg cggatcacgg ctcactacca gtactacaat gcttcggttc aatattcgtt 660aatggccgcg caggatgcgg tctgtcctgt ttgggtgcac tccgaaggaa agcaatactg 720ctcgtctact atggaacgcg cccagcagga tgttacgggt tctgagtgag tggtaatctc 780aaatttgtgt ctacatgctg gtgactaacc gctacaatgt agtgacccac gagaactccc 840tttcgatcgt gtcttcggag atccctctct gcctccagcg attttgtatg cggatatagc 900gtccccgatg ttcaaggaat ttcaccagtc actgagtacg atggcgaaag aaggacaagt 960ctcgtatcgc gtgcgataca gacctcctca acattggtct ccacgtcctg tttttgtgtc 1020tggatacggt gtcgagctgg cgttaaagcg gacggactat attgtgattg atgatagaga 1080cgcggaagaa agagggaccg gcagcattga gtccggaaag tctgatgaga cagaagatga 1140tttggatgac ctgagacccc tgtcatcatc cgaagtttct cggcttgggc tgaacacggt 1200cgggtatgtg ttggatagcg atgacccgtt tgacacactt gtgaagctgt cacaggattt 1260ccccaaatac tccgcacgtg ttgcggctca caacgtttcc accgagctgt tgcaagatgt 1320tcggtccagc agattgcgta tgcttccgcc ggggctcaac gtgctctgga tcaacggtgt 1380tcagattgaa cctcgacaag tggacgcatt cactcttctg gatcacttgc gtcgcgaaag 1440gaaattgatc gagaagttcc gaaacttagg cctgtccgct acagatgctg tagagctttt 1500gtcacaccct ctgcttggag aggccttggc acgggatggc cctcagcgtt acaactaccg 1560tgatgacatt gagggaggtg gtgtcatcat gtggctgaac aatctcgaaa aggatgcgcg 1620ctatgaatcg tggcctagcg aactcgcagg agtaggtaca tccgggattc tttattgcgg 1680agatgctaat gtctggtagt ttatgcaacg cacatatcca ggccagcttc cggcagtccg 1740ccgcgattcc aacaatattg tctttcctgt cgacttgacg agcactgaag atgctgatat 1800tgttgtcaag acaatccagg tctttgtgaa gaacaaaatt cccgtcagat ttggtttgat 1860tccggtcaca ttctcagacg gagcaattgc tcagctcaag gtcgctcatt accttcaaga 1920gacttttggt ctggccagtt ttatggatta ccttgaagcg gtaaggacta actttcccgt 1980aaccaaattc cgctattgtt tgtcagttac taattggagc agtcggcgtc caaaaataag 2040ttggcttctc cggataaggc ctgcttccag gctgcaactc aggaccggag tcctcgtctg 2100gagaaggtgt ctctatctct agatgaagtc ttgaataatg ctgtatatga cgcaacggta 2160tcaaagacaa ctgcgtacct aaaccgtctg gggatgaagc acgagccatc acatgctttt 2220gttaacggca ttcctgtcac ccgcaatgac aaatgggcgc aggaaatgag cacaaaaata 2280agcaaagata ctcagctaat tcagcagaag attgctgatg ccgaggtcga tgaagatacc 2340tggttgccag aattgtttct ctcgcaggct ttcgataggc gcaatccggc gatcgttcca 2400gaggacccga aagagatccg ggctgtggac ttggtgcagc ttgcggactc ccaagagaag 2460ctcttcagtc agattccacg tttagggcta gatgaaagca atgccttgga gagtgcccat 2520gccatcgttg ttggcaactt tgatgagaaa tccggttacg agctactcag cgcggccctt 2580gagagccgaa aaacacatgg tgaagttgag atgcttttcc tacacaatcc taagctcgag 2640gcgtcccccg catctaggtc tgtcgctgtt cgtcgattgt tgaatggtgg caaagaggta 2700gatgccagcc agattttgga ggcgatcgcc tcttccgcct cgccagcaga tgaggaagct 2760ggggatgcgg cactcttctg ggaggctcag cgagctgtag tagaagagct tggactcgct 2820ccgggcgaaa gggcacttgt catcaacgga agggtcgttg gaccgattgc agaagacacc 2880gccctgacct cagaggacct ggaccagcta ctgatatatg agaagcaaaa gcggattact 2940ccggtagcaa aggcggtcaa agcccttgaa ttcgacgaga agctttctga tccgctagac 3000tttgccaagc ttacctcgct caccacgctg tccacgatct cggatgtgcc agagggcata 3060tatgagtcga cttcggacat tcggttgaat ttgttcaaca gatggaacga ctcacaatca 3120gctatcactg tctccaattc cgatgatcca gcaattacca ttgtagcatc tatcgatccg 3180acttcggaag ttgctcagaa gtggctacca attctaaaag tactgtcgga gctggcaagt 3240gtcagagtga gattggtcct gaacccgcgc gaggagatca aagagctgcc caccaagcgc 3300ttctatcgtt atgttctcga ttcggagcca tcgttcaacg aagatgggtc ggtttcccgg 3360cccacagcct ccttctcggg cgttcccgtc gaggcactcc tcaccctggg catggatgtt 3420ccctcttctt ggcttgtggc tcccaaggat tctatccacg atcttgacaa tatcaagtta 3480agttccgtca aggacggctc gaatgtcgat gctatttacg cattggaaca catcttgatc 3540gagggccact cccgggatat gaccacgaag tccccaccta gaggagttca gcttgtcctt 3600ggaactgaga acaaccctca cttctcggat acaatcatca tggccaatct cggatacttc 3660caattcaaag cccaacctgg actgtggaac atcaacctca aaccgggccg tagcgaacgc 3720atcttcaccc tcgacagcgt aggcagcctc ggctacaacc cccaacctgg cgacgaaaac 3780aacgaagtgg ccctcctctc cttccaaggc cgcacccttt tcccgcgtgt ctcccgtaag 3840aagggctacg agaccgaaga cgtcctcgag accaacccca aaccaggttc tgcgatggac 3900tacatgaata aggggttcaa cttcgcctcc ggtatcctct ccagcgtcgg agtcggcacc 3960aaaggcagca ctagcggcaa acaggctgac attaacatct tctccgtcgc cagtggacac 4020ctctacgagc gcatgctcaa cattatgatg gtctcagtga tgcgcaacac caaccacagc 4080gtgaaattct ggttcatcga acaattcctc tccccgtcct tcaagtcctt cctgcctcac 4140cttgcgaagg agtataactt ctcttacgaa atggtcacct acaaatggcc acactggctc 4200cgggcccaga aagaaaagca acgtgaaatc tggggctaca agatcctctt cctggacgtt 4260ctcttccctc tcgacctcga caaagtcatc tttgtcgacg ccgaccagat agtccgcaca 4320gatatgtacg acctcgtcag ccttgacctc gaaggcgctc cgtacggctt tactcccatg 4380tgcgactccc gccacgagat ggaaggcttc cgcttctgga agcaggggta ctggaagaac 4440ttcctccgtg gtcaacccta ccatatctcc gcgctttacg ttgtcgacct gaaccgcttc 4500cgtgccatcg ccgccggcga tcgcctgcgt ggacagtacc agatgctgtc agctgacccc 4560gagagtttga gcaacctgga ccaggatctg ccgaaccaca tgcagcatca tatcccgatc 4620aagagtctgc cgcaggagtg gctgtggtgt gagacttggt gctcggatga gtcgcagtca 4680caggctcgga cgatcgacct gtgcaataac ccgatgacga aggagccgaa gttggatcgt 4740gccaggaggc aggtacctga gtggacggag tatgatgatg agattgcggc cttgtcgaag 4800agagttgccg ctgagaagca gcaggggcag gtggaggaag aaagggccgg tgaatcgtac 4860cctgacgagg atgaggaggg cgagacttcc tctggctggg ataaggatga gctttagcgg 4920gtttcgtttc aattatagcg tgtatacata gatcagtttg gtctccaata gggaatagat 4980tgttcgcttt acaagtcttg gtatcgtttc gtgcatgata ttcttttagt tgactgacct 5040aggatcgtaa tgccttggct tctcaatcct ataagaccta cattgggaaa cacaagcatt 5100ctcttactcg agaaaca 5117234488DNAAspergillus nigerCDS(1)..(4488) 23atg gta tct gga ctg gca aat ttc gcc tca tgg cga ctt gca tct gta 48Met Val Ser Gly Leu Ala Asn Phe Ala Ser Trp Arg Leu Ala Ser Val 1 5 10 15 ttg att gcc ggc ctg ctg gct atc cag gga cgc gct agt cca tca gtc 96Leu Ile Ala Gly Leu Leu Ala Ile Gln Gly Arg Ala Ser Pro Ser Val 20 25 30 aat gtt gct ctc caa gct tcg ttt gat tcc cca cct tat ctg ata gag 144Asn Val Ala Leu Gln Ala Ser Phe Asp Ser Pro Pro Tyr Leu Ile Glu 35

40 45 cta ctc gaa tcc gct gcg gag gag aac tcc acc tca tac ttc ccg tta 192Leu Leu Glu Ser Ala Ala Glu Glu Asn Ser Thr Ser Tyr Phe Pro Leu 50 55 60 ctc gat cgg atc gcc gac ggt att ttc gat gac gct gtt acg gat aag 240Leu Asp Arg Ile Ala Asp Gly Ile Phe Asp Asp Ala Val Thr Asp Lys 65 70 75 80 gac cta tat gat cgc ttc ctg gag gtt gtg cgt gag gat gga cac tta 288Asp Leu Tyr Asp Arg Phe Leu Glu Val Val Arg Glu Asp Gly His Leu 85 90 95 cgg acc cct gaa agt ctc tca tct ttc aag ctg tcg ctg gcg atg aga 336Arg Thr Pro Glu Ser Leu Ser Ser Phe Lys Leu Ser Leu Ala Met Arg 100 105 110 tcc gcc agt ccg cgg atc acg gct cac tac cag tac tac aat gct tcg 384Ser Ala Ser Pro Arg Ile Thr Ala His Tyr Gln Tyr Tyr Asn Ala Ser 115 120 125 gtt caa tat tcg tta atg gcc gcg cag gat gcg gtc tgt cct gtt tgg 432Val Gln Tyr Ser Leu Met Ala Ala Gln Asp Ala Val Cys Pro Val Trp 130 135 140 gtg cac tcc gaa gga aag caa tac tgc tcg tct act atg gaa cgc gcc 480Val His Ser Glu Gly Lys Gln Tyr Cys Ser Ser Thr Met Glu Arg Ala 145 150 155 160 cag cag gat gtt acg ggt tct gat gac cca cga gaa ctc cct ttc gat 528Gln Gln Asp Val Thr Gly Ser Asp Asp Pro Arg Glu Leu Pro Phe Asp 165 170 175 cgt gtc ttc gga gat ccc tct ctg cct cca gcg att ttg tat gcg gat 576Arg Val Phe Gly Asp Pro Ser Leu Pro Pro Ala Ile Leu Tyr Ala Asp 180 185 190 ata gcg tcc ccg atg ttc aag gaa ttt cac cag tca ctg agt acg atg 624Ile Ala Ser Pro Met Phe Lys Glu Phe His Gln Ser Leu Ser Thr Met 195 200 205 gcg aaa gaa gga caa gtc tcg tat cgc gtg cga tac aga cct cct caa 672Ala Lys Glu Gly Gln Val Ser Tyr Arg Val Arg Tyr Arg Pro Pro Gln 210 215 220 cat tgg tct cca cgt cct gtt ttt gtg tct gga tac ggt gtc gag ctg 720His Trp Ser Pro Arg Pro Val Phe Val Ser Gly Tyr Gly Val Glu Leu 225 230 235 240 gcg tta aag cgg acg gac tat att gtg att gat gat aga gac gcg gaa 768Ala Leu Lys Arg Thr Asp Tyr Ile Val Ile Asp Asp Arg Asp Ala Glu 245 250 255 gaa aga ggg acc ggc agc att gag tcc gga aag tct gat gag aca gaa 816Glu Arg Gly Thr Gly Ser Ile Glu Ser Gly Lys Ser Asp Glu Thr Glu 260 265 270 gat gat ttg gat gac ctg aga ccc ctg tca tca tcc gaa gtt tct cgg 864Asp Asp Leu Asp Asp Leu Arg Pro Leu Ser Ser Ser Glu Val Ser Arg 275 280 285 ctt ggg ctg aac acg gtc ggg tat gtg ttg gat agc gat gac ccg ttt 912Leu Gly Leu Asn Thr Val Gly Tyr Val Leu Asp Ser Asp Asp Pro Phe 290 295 300 gac aca ctt gtg aag ctg tca cag gat ttc ccc aaa tac tcc gca cgt 960Asp Thr Leu Val Lys Leu Ser Gln Asp Phe Pro Lys Tyr Ser Ala Arg 305 310 315 320 gtt gcg gct cac aac gtt tcc acc gag ctg ttg caa gat gtt cgg tcc 1008Val Ala Ala His Asn Val Ser Thr Glu Leu Leu Gln Asp Val Arg Ser 325 330 335 agc aga ttg cgt atg ctt ccg ccg ggg ctc aac gtg ctc tgg atc aac 1056Ser Arg Leu Arg Met Leu Pro Pro Gly Leu Asn Val Leu Trp Ile Asn 340 345 350 ggt gtt cag att gaa cct cga caa gtg gac gca ttc act ctt ctg gat 1104Gly Val Gln Ile Glu Pro Arg Gln Val Asp Ala Phe Thr Leu Leu Asp 355 360 365 cac ttg cgt cgc gaa agg aaa ttg atc gag aag ttc cga aac tta ggc 1152His Leu Arg Arg Glu Arg Lys Leu Ile Glu Lys Phe Arg Asn Leu Gly 370 375 380 ctg tcc gct aca gat gct gta gag ctt ttg tca cac cct ctg ctt gga 1200Leu Ser Ala Thr Asp Ala Val Glu Leu Leu Ser His Pro Leu Leu Gly 385 390 395 400 gag gcc ttg gca cgg gat ggc cct cag cgt tac aac tac cgt gat gac 1248Glu Ala Leu Ala Arg Asp Gly Pro Gln Arg Tyr Asn Tyr Arg Asp Asp 405 410 415 att gag gga ggt ggt gtc atc atg tgg ctg aac aat ctc gaa aag gat 1296Ile Glu Gly Gly Gly Val Ile Met Trp Leu Asn Asn Leu Glu Lys Asp 420 425 430 gcg cgc tat gaa tcg tgg cct agc gaa ctc gca gga ttt atg caa cgc 1344Ala Arg Tyr Glu Ser Trp Pro Ser Glu Leu Ala Gly Phe Met Gln Arg 435 440 445 aca tat cca ggc cag ctt ccg gca gtc cgc cgc gat tcc aac aat att 1392Thr Tyr Pro Gly Gln Leu Pro Ala Val Arg Arg Asp Ser Asn Asn Ile 450 455 460 gtc ttt cct gtc gac ttg acg agc act gaa gat gct gat att gtt gtc 1440Val Phe Pro Val Asp Leu Thr Ser Thr Glu Asp Ala Asp Ile Val Val 465 470 475 480 aag aca atc cag gtc ttt gtg aag aac aaa att ccc gtc aga ttt ggt 1488Lys Thr Ile Gln Val Phe Val Lys Asn Lys Ile Pro Val Arg Phe Gly 485 490 495 ttg att ccg gtc aca ttc tca gac gga gca att gct cag ctc aag gtc 1536Leu Ile Pro Val Thr Phe Ser Asp Gly Ala Ile Ala Gln Leu Lys Val 500 505 510 gct cat tac ctt caa gag act ttt ggt ctg gcc agt ttt atg gat tac 1584Ala His Tyr Leu Gln Glu Thr Phe Gly Leu Ala Ser Phe Met Asp Tyr 515 520 525 ctt gaa gcg tcg gcg tcc aaa aat aag ttg gct tct ccg gat aag gcc 1632Leu Glu Ala Ser Ala Ser Lys Asn Lys Leu Ala Ser Pro Asp Lys Ala 530 535 540 tgc ttc cag gct gca act cag gac cgg agt cct cgt ctg gag aag gtg 1680Cys Phe Gln Ala Ala Thr Gln Asp Arg Ser Pro Arg Leu Glu Lys Val 545 550 555 560 tct cta tct cta gat gaa gtc ttg aat aat gct gta tat gac gca acg 1728Ser Leu Ser Leu Asp Glu Val Leu Asn Asn Ala Val Tyr Asp Ala Thr 565 570 575 gta tca aag aca act gcg tac cta aac cgt ctg ggg atg aag cac gag 1776Val Ser Lys Thr Thr Ala Tyr Leu Asn Arg Leu Gly Met Lys His Glu 580 585 590 cca tca cat gct ttt gtt aac ggc att cct gtc acc cgc aat gac aaa 1824Pro Ser His Ala Phe Val Asn Gly Ile Pro Val Thr Arg Asn Asp Lys 595 600 605 tgg gcg cag gaa atg agc aca aaa ata agc aaa gat act cag cta att 1872Trp Ala Gln Glu Met Ser Thr Lys Ile Ser Lys Asp Thr Gln Leu Ile 610 615 620 cag cag aag att gct gat gcc gag gtc gat gaa gat acc tgg ttg cca 1920Gln Gln Lys Ile Ala Asp Ala Glu Val Asp Glu Asp Thr Trp Leu Pro 625 630 635 640 gaa ttg ttt ctc tcg cag gct ttc gat agg cgc aat ccg gcg atc gtt 1968Glu Leu Phe Leu Ser Gln Ala Phe Asp Arg Arg Asn Pro Ala Ile Val 645 650 655 cca gag gac ccg aaa gag atc cgg gct gtg gac ttg gtg cag ctt gcg 2016Pro Glu Asp Pro Lys Glu Ile Arg Ala Val Asp Leu Val Gln Leu Ala 660 665 670 gac tcc caa gag aag ctc ttc agt cag att cca cgt tta ggg cta gat 2064Asp Ser Gln Glu Lys Leu Phe Ser Gln Ile Pro Arg Leu Gly Leu Asp 675 680 685 gaa agc aat gcc ttg gag agt gcc cat gcc atc gtt gtt ggc aac ttt 2112Glu Ser Asn Ala Leu Glu Ser Ala His Ala Ile Val Val Gly Asn Phe 690 695 700 gat gag aaa tcc ggt tac gag cta ctc agc gcg gcc ctt gag agc cga 2160Asp Glu Lys Ser Gly Tyr Glu Leu Leu Ser Ala Ala Leu Glu Ser Arg 705 710 715 720 aaa aca cat ggt gaa gtt gag atg ctt ttc cta cac aat cct aag ctc 2208Lys Thr His Gly Glu Val Glu Met Leu Phe Leu His Asn Pro Lys Leu 725 730 735 gag gcg tcc ccc gca tct agg tct gtc gct gtt cgt cga ttg ttg aat 2256Glu Ala Ser Pro Ala Ser Arg Ser Val Ala Val Arg Arg Leu Leu Asn 740 745 750 ggt ggc aaa gag gta gat gcc agc cag att ttg gag gcg atc gcc tct 2304Gly Gly Lys Glu Val Asp Ala Ser Gln Ile Leu Glu Ala Ile Ala Ser 755 760 765 tcc gcc tcg cca gca gat gag gaa gct ggg gat gcg gca ctc ttc tgg 2352Ser Ala Ser Pro Ala Asp Glu Glu Ala Gly Asp Ala Ala Leu Phe Trp 770 775 780 gag gct cag cga gct gta gta gaa gag ctt gga ctc gct ccg ggc gaa 2400Glu Ala Gln Arg Ala Val Val Glu Glu Leu Gly Leu Ala Pro Gly Glu 785 790 795 800 agg gca ctt gtc atc aac gga agg gtc gtt gga ccg att gca gaa gac 2448Arg Ala Leu Val Ile Asn Gly Arg Val Val Gly Pro Ile Ala Glu Asp 805 810 815 acc gcc ctg acc tca gag gac ctg gac cag cta ctg ata tat gag aag 2496Thr Ala Leu Thr Ser Glu Asp Leu Asp Gln Leu Leu Ile Tyr Glu Lys 820 825 830 caa aag cgg att act ccg gta gca aag gcg gtc aaa gcc ctt gaa ttc 2544Gln Lys Arg Ile Thr Pro Val Ala Lys Ala Val Lys Ala Leu Glu Phe 835 840 845 gac gag aag ctt tct gat ccg cta gac ttt gcc aag ctt acc tcg ctc 2592Asp Glu Lys Leu Ser Asp Pro Leu Asp Phe Ala Lys Leu Thr Ser Leu 850 855 860 acc acg ctg tcc acg atc tcg gat gtg cca gag ggc ata tat gag tcg 2640Thr Thr Leu Ser Thr Ile Ser Asp Val Pro Glu Gly Ile Tyr Glu Ser 865 870 875 880 act tcg gac att cgg ttg aat ttg ttc aac aga tgg aac gac tca caa 2688Thr Ser Asp Ile Arg Leu Asn Leu Phe Asn Arg Trp Asn Asp Ser Gln 885 890 895 tca gct atc act gtc tcc aat tcc gat gat cca gca att acc att gta 2736Ser Ala Ile Thr Val Ser Asn Ser Asp Asp Pro Ala Ile Thr Ile Val 900 905 910 gca tct atc gat ccg act tcg gaa gtt gct cag aag tgg cta cca att 2784Ala Ser Ile Asp Pro Thr Ser Glu Val Ala Gln Lys Trp Leu Pro Ile 915 920 925 cta aaa gta ctg tcg gag ctg gca agt gtc aga gtg aga ttg gtc ctg 2832Leu Lys Val Leu Ser Glu Leu Ala Ser Val Arg Val Arg Leu Val Leu 930 935 940 aac ccg cgc gag gag atc aaa gag ctg ccc acc aag cgc ttc tat cgt 2880Asn Pro Arg Glu Glu Ile Lys Glu Leu Pro Thr Lys Arg Phe Tyr Arg 945 950 955 960 tat gtt ctc gat tcg gag cca tcg ttc aac gaa gat ggg tcg gtt tcc 2928Tyr Val Leu Asp Ser Glu Pro Ser Phe Asn Glu Asp Gly Ser Val Ser 965 970 975 cgg ccc aca gcc tcc ttc tcg ggc gtt ccc gtc gag gca ctc ctc acc 2976Arg Pro Thr Ala Ser Phe Ser Gly Val Pro Val Glu Ala Leu Leu Thr 980 985 990 ctg ggc atg gat gtt ccc tct tct tgg ctt gtg gct ccc aag gat tct 3024Leu Gly Met Asp Val Pro Ser Ser Trp Leu Val Ala Pro Lys Asp Ser 995 1000 1005 atc cac gat ctt gac aat atc aag tta agt tcc gtc aag gac ggc tcg 3072Ile His Asp Leu Asp Asn Ile Lys Leu Ser Ser Val Lys Asp Gly Ser 1010 1015 1020 aat gtc gat gct att tac gca ttg gaa cac atc ttg atc gag ggc cac 3120Asn Val Asp Ala Ile Tyr Ala Leu Glu His Ile Leu Ile Glu Gly His 1025 1030 1035 1040tcc cgg gat atg acc acg aag tcc cca cct aga gga gtt cag ctt gtc 3168Ser Arg Asp Met Thr Thr Lys Ser Pro Pro Arg Gly Val Gln Leu Val 1045 1050 1055 ctt gga act gag aac aac cct cac ttc tcg gat aca atc atc atg gcc 3216Leu Gly Thr Glu Asn Asn Pro His Phe Ser Asp Thr Ile Ile Met Ala 1060 1065 1070 aat ctc gga tac ttc caa ttc aaa gcc caa cct gga ctg tgg aac atc 3264Asn Leu Gly Tyr Phe Gln Phe Lys Ala Gln Pro Gly Leu Trp Asn Ile 1075 1080 1085 aac ctc aaa ccg ggc cgt agc gaa cgc atc ttc acc ctc gac agc gta 3312Asn Leu Lys Pro Gly Arg Ser Glu Arg Ile Phe Thr Leu Asp Ser Val 1090 1095 1100 ggc agc ctc ggc tac aac ccc caa cct ggc gac gaa aac aac gaa gtg 3360Gly Ser Leu Gly Tyr Asn Pro Gln Pro Gly Asp Glu Asn Asn Glu Val 1105 1110 1115 1120gcc ctc ctc tcc ttc caa ggc cgc acc ctt ttc ccg cgt gtc tcc cgt 3408Ala Leu Leu Ser Phe Gln Gly Arg Thr Leu Phe Pro Arg Val Ser Arg 1125 1130 1135 aag aag ggc tac gag acc gaa gac gtc ctc gag acc aac ccc aaa cca 3456Lys Lys Gly Tyr Glu Thr Glu Asp Val Leu Glu Thr Asn Pro Lys Pro 1140 1145 1150 ggt tct gcg atg gac tac atg aat aag ggg ttc aac ttc gcc tcc ggt 3504Gly Ser Ala Met Asp Tyr Met Asn Lys Gly Phe Asn Phe Ala Ser Gly 1155 1160 1165 atc ctc tcc agc gtc gga gtc ggc acc aaa ggc agc act agc ggc aaa 3552Ile Leu Ser Ser Val Gly Val Gly Thr Lys Gly Ser Thr Ser Gly Lys 1170 1175 1180 cag gct gac att aac atc ttc tcc gtc gcc agt gga cac ctc tac gag 3600Gln Ala Asp Ile Asn Ile Phe Ser Val Ala Ser Gly His Leu Tyr Glu 1185 1190 1195 1200cgc atg ctc aac att atg atg gtc tca gtg atg cgc aac acc aac cac 3648Arg Met Leu Asn Ile Met Met Val Ser Val Met Arg Asn Thr Asn His 1205 1210 1215 agc gtg aaa ttc tgg ttc atc gaa caa ttc ctc tcc ccg tcc ttc aag 3696Ser Val Lys Phe Trp Phe Ile Glu Gln Phe Leu Ser Pro Ser Phe Lys 1220 1225 1230 tcc ttc ctg cct cac ctt gcg aag gag tat aac ttc tct tac gaa atg 3744Ser Phe Leu Pro His Leu Ala Lys Glu Tyr Asn Phe Ser Tyr Glu Met 1235 1240 1245 gtc acc tac aaa tgg cca cac tgg ctc cgg gcc cag aaa gaa aag caa 3792Val Thr Tyr Lys Trp Pro His Trp Leu Arg Ala Gln Lys Glu Lys Gln 1250 1255 1260 cgt gaa atc tgg ggc tac aag atc ctc ttc ctg gac gtt ctc ttc cct 3840Arg Glu Ile Trp Gly Tyr Lys Ile Leu Phe Leu Asp Val Leu Phe Pro 1265 1270 1275 1280ctc gac ctc gac aaa gtc atc ttt gtc gac gcc gac cag ata gtc cgc 3888Leu Asp Leu Asp Lys Val Ile Phe Val Asp Ala Asp Gln Ile Val Arg 1285 1290 1295 aca gat atg tac gac ctc gtc agc ctt gac ctc gaa ggc gct ccg tac 3936Thr Asp Met Tyr Asp Leu Val Ser Leu Asp Leu Glu Gly Ala Pro Tyr 1300 1305 1310 ggc ttt act ccc atg tgc gac tcc cgc cac gag atg gaa ggc ttc cgc 3984Gly Phe Thr Pro Met Cys Asp Ser Arg His Glu Met Glu Gly Phe Arg 1315 1320 1325 ttc tgg aag cag ggg tac tgg aag aac ttc ctc cgt ggt caa ccc tac 4032Phe Trp Lys Gln Gly Tyr Trp Lys Asn Phe Leu Arg Gly Gln Pro Tyr 1330 1335 1340 cat atc tcc gcg ctt tac gtt gtc gac ctg aac cgc ttc cgt gcc atc 4080His Ile Ser Ala Leu Tyr Val Val Asp Leu Asn Arg Phe Arg Ala Ile 1345 1350 1355 1360gcc gcc ggc gat cgc ctg cgt gga cag tac cag atg ctg tca gct gac 4128Ala Ala Gly Asp Arg Leu Arg Gly Gln Tyr Gln Met Leu Ser Ala Asp 1365 1370 1375 ccc gag agt ttg agc aac ctg gac cag gat ctg ccg aac cac atg cag 4176Pro Glu Ser Leu Ser Asn Leu Asp Gln Asp Leu Pro Asn His Met Gln 1380 1385 1390 cat cat atc ccg atc aag agt ctg ccg cag gag tgg ctg tgg tgt gag 4224His His Ile Pro Ile Lys Ser Leu Pro Gln Glu Trp Leu Trp Cys Glu 1395 1400 1405 act tgg tgc tcg gat gag tcg cag tca cag gct cgg acg atc gac ctg 4272Thr Trp Cys Ser Asp Glu Ser Gln Ser Gln Ala Arg Thr Ile Asp Leu 1410 1415 1420 tgc aat aac ccg atg acg aag gag ccg aag ttg gat cgt gcc agg agg 4320Cys Asn Asn Pro Met Thr Lys Glu Pro Lys Leu Asp Arg Ala Arg Arg 1425 1430 1435 1440cag gta cct gag tgg acg gag tat gat gat gag att gcg gcc ttg tcg 4368Gln Val Pro Glu Trp Thr Glu Tyr Asp Asp Glu Ile Ala Ala Leu Ser 1445 1450 1455 aag aga gtt gcc gct gag aag cag cag ggg cag gtg gag gaa gaa agg 4416Lys Arg Val Ala Ala Glu Lys Gln Gln Gly Gln Val Glu Glu Glu Arg 1460 1465 1470 gcc ggt gaa tcg tac cct gac gag gat gag gag ggc gag act tcc tct 4464Ala Gly Glu Ser Tyr Pro Asp Glu Asp Glu Glu Gly Glu Thr Ser Ser 1475 1480 1485 ggc tgg gat aag gat gag ctt tag 4488Gly Trp Asp Lys Asp Glu Leu 1490 1495241495PRTAspergillus niger 24Met Val Ser Gly Leu Ala Asn Phe Ala Ser

Trp Arg Leu Ala Ser Val 1 5 10 15 Leu Ile Ala Gly Leu Leu Ala Ile Gln Gly Arg Ala Ser Pro Ser Val 20 25 30 Asn Val Ala Leu Gln Ala Ser Phe Asp Ser Pro Pro Tyr Leu Ile Glu 35 40 45 Leu Leu Glu Ser Ala Ala Glu Glu Asn Ser Thr Ser Tyr Phe Pro Leu 50 55 60 Leu Asp Arg Ile Ala Asp Gly Ile Phe Asp Asp Ala Val Thr Asp Lys 65 70 75 80 Asp Leu Tyr Asp Arg Phe Leu Glu Val Val Arg Glu Asp Gly His Leu 85 90 95 Arg Thr Pro Glu Ser Leu Ser Ser Phe Lys Leu Ser Leu Ala Met Arg 100 105 110 Ser Ala Ser Pro Arg Ile Thr Ala His Tyr Gln Tyr Tyr Asn Ala Ser 115 120 125 Val Gln Tyr Ser Leu Met Ala Ala Gln Asp Ala Val Cys Pro Val Trp 130 135 140 Val His Ser Glu Gly Lys Gln Tyr Cys Ser Ser Thr Met Glu Arg Ala 145 150 155 160 Gln Gln Asp Val Thr Gly Ser Asp Asp Pro Arg Glu Leu Pro Phe Asp 165 170 175 Arg Val Phe Gly Asp Pro Ser Leu Pro Pro Ala Ile Leu Tyr Ala Asp 180 185 190 Ile Ala Ser Pro Met Phe Lys Glu Phe His Gln Ser Leu Ser Thr Met 195 200 205 Ala Lys Glu Gly Gln Val Ser Tyr Arg Val Arg Tyr Arg Pro Pro Gln 210 215 220 His Trp Ser Pro Arg Pro Val Phe Val Ser Gly Tyr Gly Val Glu Leu 225 230 235 240 Ala Leu Lys Arg Thr Asp Tyr Ile Val Ile Asp Asp Arg Asp Ala Glu 245 250 255 Glu Arg Gly Thr Gly Ser Ile Glu Ser Gly Lys Ser Asp Glu Thr Glu 260 265 270 Asp Asp Leu Asp Asp Leu Arg Pro Leu Ser Ser Ser Glu Val Ser Arg 275 280 285 Leu Gly Leu Asn Thr Val Gly Tyr Val Leu Asp Ser Asp Asp Pro Phe 290 295 300 Asp Thr Leu Val Lys Leu Ser Gln Asp Phe Pro Lys Tyr Ser Ala Arg 305 310 315 320 Val Ala Ala His Asn Val Ser Thr Glu Leu Leu Gln Asp Val Arg Ser 325 330 335 Ser Arg Leu Arg Met Leu Pro Pro Gly Leu Asn Val Leu Trp Ile Asn 340 345 350 Gly Val Gln Ile Glu Pro Arg Gln Val Asp Ala Phe Thr Leu Leu Asp 355 360 365 His Leu Arg Arg Glu Arg Lys Leu Ile Glu Lys Phe Arg Asn Leu Gly 370 375 380 Leu Ser Ala Thr Asp Ala Val Glu Leu Leu Ser His Pro Leu Leu Gly 385 390 395 400 Glu Ala Leu Ala Arg Asp Gly Pro Gln Arg Tyr Asn Tyr Arg Asp Asp 405 410 415 Ile Glu Gly Gly Gly Val Ile Met Trp Leu Asn Asn Leu Glu Lys Asp 420 425 430 Ala Arg Tyr Glu Ser Trp Pro Ser Glu Leu Ala Gly Phe Met Gln Arg 435 440 445 Thr Tyr Pro Gly Gln Leu Pro Ala Val Arg Arg Asp Ser Asn Asn Ile 450 455 460 Val Phe Pro Val Asp Leu Thr Ser Thr Glu Asp Ala Asp Ile Val Val 465 470 475 480 Lys Thr Ile Gln Val Phe Val Lys Asn Lys Ile Pro Val Arg Phe Gly 485 490 495 Leu Ile Pro Val Thr Phe Ser Asp Gly Ala Ile Ala Gln Leu Lys Val 500 505 510 Ala His Tyr Leu Gln Glu Thr Phe Gly Leu Ala Ser Phe Met Asp Tyr 515 520 525 Leu Glu Ala Ser Ala Ser Lys Asn Lys Leu Ala Ser Pro Asp Lys Ala 530 535 540 Cys Phe Gln Ala Ala Thr Gln Asp Arg Ser Pro Arg Leu Glu Lys Val 545 550 555 560 Ser Leu Ser Leu Asp Glu Val Leu Asn Asn Ala Val Tyr Asp Ala Thr 565 570 575 Val Ser Lys Thr Thr Ala Tyr Leu Asn Arg Leu Gly Met Lys His Glu 580 585 590 Pro Ser His Ala Phe Val Asn Gly Ile Pro Val Thr Arg Asn Asp Lys 595 600 605 Trp Ala Gln Glu Met Ser Thr Lys Ile Ser Lys Asp Thr Gln Leu Ile 610 615 620 Gln Gln Lys Ile Ala Asp Ala Glu Val Asp Glu Asp Thr Trp Leu Pro 625 630 635 640 Glu Leu Phe Leu Ser Gln Ala Phe Asp Arg Arg Asn Pro Ala Ile Val 645 650 655 Pro Glu Asp Pro Lys Glu Ile Arg Ala Val Asp Leu Val Gln Leu Ala 660 665 670 Asp Ser Gln Glu Lys Leu Phe Ser Gln Ile Pro Arg Leu Gly Leu Asp 675 680 685 Glu Ser Asn Ala Leu Glu Ser Ala His Ala Ile Val Val Gly Asn Phe 690 695 700 Asp Glu Lys Ser Gly Tyr Glu Leu Leu Ser Ala Ala Leu Glu Ser Arg 705 710 715 720 Lys Thr His Gly Glu Val Glu Met Leu Phe Leu His Asn Pro Lys Leu 725 730 735 Glu Ala Ser Pro Ala Ser Arg Ser Val Ala Val Arg Arg Leu Leu Asn 740 745 750 Gly Gly Lys Glu Val Asp Ala Ser Gln Ile Leu Glu Ala Ile Ala Ser 755 760 765 Ser Ala Ser Pro Ala Asp Glu Glu Ala Gly Asp Ala Ala Leu Phe Trp 770 775 780 Glu Ala Gln Arg Ala Val Val Glu Glu Leu Gly Leu Ala Pro Gly Glu 785 790 795 800 Arg Ala Leu Val Ile Asn Gly Arg Val Val Gly Pro Ile Ala Glu Asp 805 810 815 Thr Ala Leu Thr Ser Glu Asp Leu Asp Gln Leu Leu Ile Tyr Glu Lys 820 825 830 Gln Lys Arg Ile Thr Pro Val Ala Lys Ala Val Lys Ala Leu Glu Phe 835 840 845 Asp Glu Lys Leu Ser Asp Pro Leu Asp Phe Ala Lys Leu Thr Ser Leu 850 855 860 Thr Thr Leu Ser Thr Ile Ser Asp Val Pro Glu Gly Ile Tyr Glu Ser 865 870 875 880 Thr Ser Asp Ile Arg Leu Asn Leu Phe Asn Arg Trp Asn Asp Ser Gln 885 890 895 Ser Ala Ile Thr Val Ser Asn Ser Asp Asp Pro Ala Ile Thr Ile Val 900 905 910 Ala Ser Ile Asp Pro Thr Ser Glu Val Ala Gln Lys Trp Leu Pro Ile 915 920 925 Leu Lys Val Leu Ser Glu Leu Ala Ser Val Arg Val Arg Leu Val Leu 930 935 940 Asn Pro Arg Glu Glu Ile Lys Glu Leu Pro Thr Lys Arg Phe Tyr Arg 945 950 955 960 Tyr Val Leu Asp Ser Glu Pro Ser Phe Asn Glu Asp Gly Ser Val Ser 965 970 975 Arg Pro Thr Ala Ser Phe Ser Gly Val Pro Val Glu Ala Leu Leu Thr 980 985 990 Leu Gly Met Asp Val Pro Ser Ser Trp Leu Val Ala Pro Lys Asp Ser 995 1000 1005 Ile His Asp Leu Asp Asn Ile Lys Leu Ser Ser Val Lys Asp Gly Ser 1010 1015 1020 Asn Val Asp Ala Ile Tyr Ala Leu Glu His Ile Leu Ile Glu Gly His 1025 1030 1035 1040Ser Arg Asp Met Thr Thr Lys Ser Pro Pro Arg Gly Val Gln Leu Val 1045 1050 1055 Leu Gly Thr Glu Asn Asn Pro His Phe Ser Asp Thr Ile Ile Met Ala 1060 1065 1070 Asn Leu Gly Tyr Phe Gln Phe Lys Ala Gln Pro Gly Leu Trp Asn Ile 1075 1080 1085 Asn Leu Lys Pro Gly Arg Ser Glu Arg Ile Phe Thr Leu Asp Ser Val 1090 1095 1100 Gly Ser Leu Gly Tyr Asn Pro Gln Pro Gly Asp Glu Asn Asn Glu Val 1105 1110 1115 1120Ala Leu Leu Ser Phe Gln Gly Arg Thr Leu Phe Pro Arg Val Ser Arg 1125 1130 1135 Lys Lys Gly Tyr Glu Thr Glu Asp Val Leu Glu Thr Asn Pro Lys Pro 1140 1145 1150 Gly Ser Ala Met Asp Tyr Met Asn Lys Gly Phe Asn Phe Ala Ser Gly 1155 1160 1165 Ile Leu Ser Ser Val Gly Val Gly Thr Lys Gly Ser Thr Ser Gly Lys 1170 1175 1180 Gln Ala Asp Ile Asn Ile Phe Ser Val Ala Ser Gly His Leu Tyr Glu 1185 1190 1195 1200Arg Met Leu Asn Ile Met Met Val Ser Val Met Arg Asn Thr Asn His 1205 1210 1215 Ser Val Lys Phe Trp Phe Ile Glu Gln Phe Leu Ser Pro Ser Phe Lys 1220 1225 1230 Ser Phe Leu Pro His Leu Ala Lys Glu Tyr Asn Phe Ser Tyr Glu Met 1235 1240 1245 Val Thr Tyr Lys Trp Pro His Trp Leu Arg Ala Gln Lys Glu Lys Gln 1250 1255 1260 Arg Glu Ile Trp Gly Tyr Lys Ile Leu Phe Leu Asp Val Leu Phe Pro 1265 1270 1275 1280Leu Asp Leu Asp Lys Val Ile Phe Val Asp Ala Asp Gln Ile Val Arg 1285 1290 1295 Thr Asp Met Tyr Asp Leu Val Ser Leu Asp Leu Glu Gly Ala Pro Tyr 1300 1305 1310 Gly Phe Thr Pro Met Cys Asp Ser Arg His Glu Met Glu Gly Phe Arg 1315 1320 1325 Phe Trp Lys Gln Gly Tyr Trp Lys Asn Phe Leu Arg Gly Gln Pro Tyr 1330 1335 1340 His Ile Ser Ala Leu Tyr Val Val Asp Leu Asn Arg Phe Arg Ala Ile 1345 1350 1355 1360Ala Ala Gly Asp Arg Leu Arg Gly Gln Tyr Gln Met Leu Ser Ala Asp 1365 1370 1375 Pro Glu Ser Leu Ser Asn Leu Asp Gln Asp Leu Pro Asn His Met Gln 1380 1385 1390 His His Ile Pro Ile Lys Ser Leu Pro Gln Glu Trp Leu Trp Cys Glu 1395 1400 1405 Thr Trp Cys Ser Asp Glu Ser Gln Ser Gln Ala Arg Thr Ile Asp Leu 1410 1415 1420 Cys Asn Asn Pro Met Thr Lys Glu Pro Lys Leu Asp Arg Ala Arg Arg 1425 1430 1435 1440Gln Val Pro Glu Trp Thr Glu Tyr Asp Asp Glu Ile Ala Ala Leu Ser 1445 1450 1455 Lys Arg Val Ala Ala Glu Lys Gln Gln Gly Gln Val Glu Glu Glu Arg 1460 1465 1470 Ala Gly Glu Ser Tyr Pro Asp Glu Asp Glu Glu Gly Glu Thr Ser Ser 1475 1480 1485 Gly Trp Asp Lys Asp Glu Leu 1490 1495251531DNAAspergillus niger 25attgtcatac agtggccgac aacctacgtt acccgaccgg gtctctgggt tgattaactg 60agcacggtcc acacgaggac cattgaggac catctctcgc gaacttactg ggctatcttg 120atggtactct agaagtgggt tgcaggacaa ttccacagtg aaagctgcgt gtcaagcttt 180ctatatatac actattgacc atgctgaacc tcaatatctt ccggctactg gccgatctct 240cccatatctc ctccaaatgt gtcttgatat gggctatcca tcgcaataag agcgcagaag 300gtccgtataa taacttgcat tgttgcagtg tgctgcgagc taagcatcgt ttcaaataat 360aggagtctcc cttctgacgc agatgctcta tgctttggtg ttcgtgactc gttatctcga 420ccttttctcg aaggcaggat ggaagcactt ctacctcgta ttcttcaagc tattttatat 480catctcctcg ttctacgtta tatacctgat gatgagagta tttccccgga cacgggaaag 540ggagcgagcc tggaagatgg ctataatatc ggtcgctcta tctctggttc tggctcctat 600atctattgtc atcttctatc gtggttatcc cgatagatgg ttcacggagg taagtgggat 660ggctcgcatt ggcctgcaga cgtcgctaac caaaccgtga tgatatgcag acttgctgga 720ctttctcgat tatattagag tccgtctgtg ttctccctca attgttgctc ttgcgccaaa 780cgaccgttcc gacagtcatc gattcatact acctgcttat gctgggatcc taccgtgcct 840tctatattct caattggctt gtgcggggac tgggctctga gggtcattgg gacgtaattg 900cagacctcta cggtgtcatc cagacggctt tctacgtcga tttcgcctgg gtttactact 960cccgccaacg cgtgaagctc cgaaacggcg gtgtcgttga ctcggaagat ttccgccata 1020gctggctagt gagcaagata ctgaatttcc ggcagcgaag gagtgcagat gaggagcaga 1080atttgaacga cgaggacgtg gaggatgagg aagttgctgg tggcggtaga cccaggaaca 1140accgctgggg agcaatgggg atctccgtct cggccgacga tacgctagga aaccatcgtg 1200ggacaagcca agacgagagt ctggaagggt tcttagaaga tgaagaagac gacgaggaca 1260ataacgggta ccctgtgaac gggggcgttc gtccgaagca gtcaaccggg gtaacgggca 1320gtcacgaatg aactatctgc ccttaaaccc catatataga aatcctgctg cagatcagcc 1380ggttttggtt acacgattca actgccctcg gggcattata tacatcccta gggctcgttc 1440tccccctttc gcttccttca tggtctgttt ctttattgct cgggtctttg tttgcatgga 1500tttctctcac gtcatcaagt tttcacaatc t 1531261008DNAAspergillus nigerCDS(1)..(1008) 26atg ctg aac ctc aat atc ttc cgg cta ctg gcc gat ctc tcc cat atc 48Met Leu Asn Leu Asn Ile Phe Arg Leu Leu Ala Asp Leu Ser His Ile 1 5 10 15 tcc tcc aaa tgt gtc ttg ata tgg gct atc cat cgc aat aag agc gca 96Ser Ser Lys Cys Val Leu Ile Trp Ala Ile His Arg Asn Lys Ser Ala 20 25 30 gaa gga gtc tcc ctt ctg acg cag atg ctc tat gct ttg gtg ttc gtg 144Glu Gly Val Ser Leu Leu Thr Gln Met Leu Tyr Ala Leu Val Phe Val 35 40 45 act cgt tat ctc gac ctt ttc tcg aag gca gga tgg aag cac ttc tac 192Thr Arg Tyr Leu Asp Leu Phe Ser Lys Ala Gly Trp Lys His Phe Tyr 50 55 60 ctc gta ttc ttc aag cta ttt tat atc atc tcc tcg ttc tac gtt ata 240Leu Val Phe Phe Lys Leu Phe Tyr Ile Ile Ser Ser Phe Tyr Val Ile 65 70 75 80 tac ctg atg atg aga gta ttt ccc cgg aca cgg gaa agg gag cga gcc 288Tyr Leu Met Met Arg Val Phe Pro Arg Thr Arg Glu Arg Glu Arg Ala 85 90 95 tgg aag atg gct ata ata tcg gtc gct cta tct ctg gtt ctg gct cct 336Trp Lys Met Ala Ile Ile Ser Val Ala Leu Ser Leu Val Leu Ala Pro 100 105 110 ata tct att gtc atc ttc tat cgt ggt tat ccc gat aga tgg ttc acg 384Ile Ser Ile Val Ile Phe Tyr Arg Gly Tyr Pro Asp Arg Trp Phe Thr 115 120 125 gag act tgc tgg act ttc tcg att ata tta gag tcc gtc tgt gtt ctc 432Glu Thr Cys Trp Thr Phe Ser Ile Ile Leu Glu Ser Val Cys Val Leu 130 135 140 cct caa ttg ttg ctc ttg cgc caa acg acc gtt ccg aca gtc atc gat 480Pro Gln Leu Leu Leu Leu Arg Gln Thr Thr Val Pro Thr Val Ile Asp 145 150 155 160 tca tac tac ctg ctt atg ctg gga tcc tac cgt gcc ttc tat att ctc 528Ser Tyr Tyr Leu Leu Met Leu Gly Ser Tyr Arg Ala Phe Tyr Ile Leu 165 170 175 aat tgg ctt gtg cgg gga ctg ggc tct gag ggt cat tgg gac gta att 576Asn Trp Leu Val Arg Gly Leu Gly Ser Glu Gly His Trp Asp Val Ile 180 185 190 gca gac ctc tac ggt gtc atc cag acg gct ttc tac gtc gat ttc gcc 624Ala Asp Leu Tyr Gly Val Ile Gln Thr Ala Phe Tyr Val Asp Phe Ala 195 200 205 tgg gtt tac tac tcc cgc caa cgc gtg aag ctc cga aac ggc ggt gtc 672Trp Val Tyr Tyr Ser Arg Gln Arg Val Lys Leu Arg Asn Gly Gly Val 210 215 220 gtt gac tcg gaa gat ttc cgc cat agc tgg cta gtg agc aag ata ctg 720Val Asp Ser Glu Asp Phe Arg His Ser Trp Leu Val Ser Lys Ile Leu 225 230 235 240 aat ttc cgg cag cga agg agt gca gat gag gag cag aat ttg aac gac 768Asn Phe Arg Gln Arg Arg Ser Ala Asp Glu Glu Gln Asn Leu Asn Asp 245 250 255 gag gac gtg gag gat gag gaa gtt gct ggt ggc ggt aga ccc agg aac 816Glu Asp Val Glu Asp Glu Glu Val Ala Gly Gly Gly Arg Pro Arg Asn 260 265 270 aac cgc tgg gga gca atg ggg atc tcc gtc tcg gcc gac gat acg cta 864Asn Arg Trp Gly Ala Met Gly Ile Ser Val Ser Ala Asp Asp Thr Leu 275 280 285 gga aac cat cgt ggg aca agc caa gac gag agt ctg gaa ggg ttc tta 912Gly Asn His Arg Gly Thr Ser Gln Asp Glu Ser Leu Glu Gly Phe Leu 290 295 300 gaa gat gaa gaa gac gac gag gac aat aac ggg tac cct gtg aac ggg 960Glu Asp Glu Glu Asp Asp Glu Asp Asn Asn Gly Tyr Pro Val Asn Gly 305 310 315 320 ggc gtt cgt ccg aag cag tca acc ggg gta acg ggc agt cac gaa tga 1008Gly Val Arg Pro Lys Gln Ser Thr Gly Val Thr Gly Ser His Glu 325 330 335 27335PRTAspergillus niger 27Met Leu Asn Leu Asn Ile Phe Arg Leu Leu Ala Asp Leu Ser His Ile 1 5 10 15 Ser Ser Lys Cys Val Leu Ile Trp Ala Ile His Arg Asn Lys Ser Ala 20 25 30 Glu

Gly Val Ser Leu Leu Thr Gln Met Leu Tyr Ala Leu Val Phe Val 35 40 45 Thr Arg Tyr Leu Asp Leu Phe Ser Lys Ala Gly Trp Lys His Phe Tyr 50 55 60 Leu Val Phe Phe Lys Leu Phe Tyr Ile Ile Ser Ser Phe Tyr Val Ile 65 70 75 80 Tyr Leu Met Met Arg Val Phe Pro Arg Thr Arg Glu Arg Glu Arg Ala 85 90 95 Trp Lys Met Ala Ile Ile Ser Val Ala Leu Ser Leu Val Leu Ala Pro 100 105 110 Ile Ser Ile Val Ile Phe Tyr Arg Gly Tyr Pro Asp Arg Trp Phe Thr 115 120 125 Glu Thr Cys Trp Thr Phe Ser Ile Ile Leu Glu Ser Val Cys Val Leu 130 135 140 Pro Gln Leu Leu Leu Leu Arg Gln Thr Thr Val Pro Thr Val Ile Asp 145 150 155 160 Ser Tyr Tyr Leu Leu Met Leu Gly Ser Tyr Arg Ala Phe Tyr Ile Leu 165 170 175 Asn Trp Leu Val Arg Gly Leu Gly Ser Glu Gly His Trp Asp Val Ile 180 185 190 Ala Asp Leu Tyr Gly Val Ile Gln Thr Ala Phe Tyr Val Asp Phe Ala 195 200 205 Trp Val Tyr Tyr Ser Arg Gln Arg Val Lys Leu Arg Asn Gly Gly Val 210 215 220 Val Asp Ser Glu Asp Phe Arg His Ser Trp Leu Val Ser Lys Ile Leu 225 230 235 240 Asn Phe Arg Gln Arg Arg Ser Ala Asp Glu Glu Gln Asn Leu Asn Asp 245 250 255 Glu Asp Val Glu Asp Glu Glu Val Ala Gly Gly Gly Arg Pro Arg Asn 260 265 270 Asn Arg Trp Gly Ala Met Gly Ile Ser Val Ser Ala Asp Asp Thr Leu 275 280 285 Gly Asn His Arg Gly Thr Ser Gln Asp Glu Ser Leu Glu Gly Phe Leu 290 295 300 Glu Asp Glu Glu Asp Asp Glu Asp Asn Asn Gly Tyr Pro Val Asn Gly 305 310 315 320 Gly Val Arg Pro Lys Gln Ser Thr Gly Val Thr Gly Ser His Glu 325 330 335 283392DNAAspergillus niger 28gcccttgttg tactactttc attatcgtat ctagttgcat tttccttctt ctatcccagc 60acataacagc tttgtgtgtg ggaccttcac ctctggtaga tgcaggtcga cacactagtg 120gttgctgata gttcttcttt cagaggttga gtgtctctga ttgactactg agctctccca 180tcatggccgg aactcggcca atgtccaacc gttggaccct actgctgtcc ttggtgatcc 240tactcggatg ccttgtcatc cccggaggta agcctatcca atcacactcc tactggtgag 300gctctccttg acatctaaca atgacgcata gtcaccgtga aacacgagaa cttcaagaca 360tgttctcaat cgggcttctg taagcggaac agagcattcg ccgacgatgc tgccgcccaa 420ggttcctcct gggcctcccc atacgaactc gactcatcct ccatccagtt caaggatggc 480caattgcacg gaaccattct caagtccgtc tcccccaacg agaaagtcaa gctgcctctc 540gttgtctcct tcctcgagtc cggcgccgcc cgagttgttg tcgatgagga aaagcgcatg 600aacggtgaca tccagcttcg acacgatagc aaagcacgca aggaacgcta caatgaggca 660gagaaatggg tgttggttgg tggcctggag ttgagcaaaa ccgcgacctt gagacctgaa 720accgagtctg gctttaccag agtcttgtac ggtccggaca accagttcga ggctgtcatc 780cgccacgccc cctttagcgc cgacttcaag agggatggcc aaacccacgt tcaattgaac 840aacaagggct accttaacat ggagcattgg cgccctaagg tagaggtcga aggcgagggc 900gagcagcaaa cccaggaaga tgaaagcact tggtgggatg agagctttgg tggaaacacg 960gacaccaagc ccaggggtcc cgagagtgtg ggattggata tcaccttccc tggctacaag 1020catgtttttg gaattcctga gcatgctgac tctctctcct taaaggaaac tcggtaagct 1080agtcgcgcag tgacattttc catctcgcag aactgacagc gtcccagagg tggtgaaggg 1140aatcacgaag agccctaccg catgtacaat gcggatgtat ttgagtacga gctgagcagt 1200cccatgacct tgtatggtgc tattccattc atgcaggcac atcgcaagga ctccaccgtc 1260ggtgtcttct ggctgaatgc tgcagagacc tgggtggaca ttgtcaagtc tacctcatct 1320cctaaccctc ttgctctcgg cgtgggcgcc accactgaca cccagagtca ttggttttcg 1380gagtccggcc agctcgacgt gttcgttttc cttggtccta ccccacagga aatcagcaag 1440acctatggtg aactcaccgg ctacactcag ttgcctcaac attttgccat tgcttatcac 1500cagtgccgct ggaactacat cactgatgag gatgtcaagg aggtcgatcg caactttgac 1560aagtaccaga tcccctacga tgtcatctgg ctggacatcg aatataccga tgacagaaag 1620tatttcacct gggatccact cagtttcccc gatccgatca gcatggagga gcagctcgat 1680gagtcggagc gcaaactcgt cgttatcatt gacccgcaca tcaagaacca ggacaagtac 1740agcatcgtcc aagaaatgaa gagcaaagac ttggccacta agaacaagga cggtgagatc 1800tacgacgggt ggtgttggcc tggctcttct cactggatcg ataccttcaa ccccgccgcc 1860atcaaatggt gggtcagctt attcaagttt gacaagttca aggggacgct gtccaatgtc 1920ttcatttgga acgacatgaa cgagccctcg gttttcaacg gtcccgaaac cacgatgccc 1980aaggataacc ttcatcatgg caactgggag caccgtgaca tccataacgt tcatggaatc 2040accctggtca atgccaccta cgatgccctt ctagagcgca agaagggcga gatccgtcgg 2100cctttcattc tgacacggtc atattatgct ggtgctcaac ggatgtctgc tatgtggacg 2160ggtgataacc aggctacttg ggaacacttg gccgcttcca tccctatggt tctgaacaac 2220ggcattgcgg gcttcccctt tgccggtgct gacgtgggcg gtttcttcca gaaccctagc 2280aaggagctct tgaccagatg gtaccaagct ggtatttggt accccttctt ccgggcccac 2340gcgcatattg acacgcgccg gagagagccg tatctgattg ccgagccaca ccggtctatc 2400atctcccagg ctatccgcct gaggtatcag cttctccccg cctggtacac tgccttccac 2460gaagcttccg tgaacggaat gccgatcgtg aggccgcagt actacgctca cccttgggat 2520gaggctggct ttgccattga cgaccagctt tatctcggct ccaccggtct tcttgctaag 2580cctgttgtct ccgaggaggc caccacggcc gacatttacc ttgctgacga cgaaaagtac 2640tatgactact ttgactacac cgtctaccag ggagccggaa agcggcatac ggtgcctgct 2700cctatggaga ctgtgccatt gctgatgcag ggtggccatg taatcccccg caaggaccgt 2760cctcgccgca gtagcgcctt gatgagatgg gatccgtaca ctcttgttgt ggtcttggat 2820aagaacggtc aagccgatgg ctctctctac gtggatgacg gtgagacgtt cgactatgag 2880cgtggagctt atatccaccg ccgtttccgc ttccaggagt ctgccctggt ctcggaggat 2940gttggcacca agggtcctaa gacggccgag tacttgaaga ccatggccaa cgttcgtgtt 3000gagcgggtgg tggtagttga tcctcctaag gaatggcagg gtaagaccag tgtgactgtc 3060attgaggatg gagcttcggc ggcttcgaca gcctctatgc agtaccacag ccagcccgat 3120ggcaaggccg catatgcggt ggtgaagaac cccaatgtcg gcattggaaa gacatggcgg 3180attgagtttt agactagacg aggatatgga ttgagcaccc atacatatat gcaagaggct 3240catatatcaa acatcaatgg tatttagtta tgacgctttc agatgccctt acaccctagt 3300tgagcaccac ccgtagtaga atcgtagtga agggtggaac cccaaccctg aagaggaaaa 3360agggaaggca actcccggag tggggctgag tc 3392292874DNAAspergillus nigerCDS(1)..(2874) 29atg tcc aac cgt tgg acc cta ctg ctg tcc ttg gtg atc cta ctc gga 48Met Ser Asn Arg Trp Thr Leu Leu Leu Ser Leu Val Ile Leu Leu Gly 1 5 10 15 tgc ctt gtc atc ccc gga gtc acc gtg aaa cac gag aac ttc aag aca 96Cys Leu Val Ile Pro Gly Val Thr Val Lys His Glu Asn Phe Lys Thr 20 25 30 tgt tct caa tcg ggc ttc tgt aag cgg aac aga gca ttc gcc gac gat 144Cys Ser Gln Ser Gly Phe Cys Lys Arg Asn Arg Ala Phe Ala Asp Asp 35 40 45 gct gcc gcc caa ggt tcc tcc tgg gcc tcc cca tac gaa ctc gac tca 192Ala Ala Ala Gln Gly Ser Ser Trp Ala Ser Pro Tyr Glu Leu Asp Ser 50 55 60 tcc tcc atc cag ttc aag gat ggc caa ttg cac gga acc att ctc aag 240Ser Ser Ile Gln Phe Lys Asp Gly Gln Leu His Gly Thr Ile Leu Lys 65 70 75 80 tcc gtc tcc ccc aac gag aaa gtc aag ctg cct ctc gtt gtc tcc ttc 288Ser Val Ser Pro Asn Glu Lys Val Lys Leu Pro Leu Val Val Ser Phe 85 90 95 ctc gag tcc ggc gcc gcc cga gtt gtt gtc gat gag gaa aag cgc atg 336Leu Glu Ser Gly Ala Ala Arg Val Val Val Asp Glu Glu Lys Arg Met 100 105 110 aac ggt gac atc cag ctt cga cac gat agc aaa gca cgc aag gaa cgc 384Asn Gly Asp Ile Gln Leu Arg His Asp Ser Lys Ala Arg Lys Glu Arg 115 120 125 tac aat gag gca gag aaa tgg gtg ttg gtt ggt ggc ctg gag ttg agc 432Tyr Asn Glu Ala Glu Lys Trp Val Leu Val Gly Gly Leu Glu Leu Ser 130 135 140 aaa acc gcg acc ttg aga cct gaa acc gag tct ggc ttt acc aga gtc 480Lys Thr Ala Thr Leu Arg Pro Glu Thr Glu Ser Gly Phe Thr Arg Val 145 150 155 160 ttg tac ggt ccg gac aac cag ttc gag gct gtc atc cgc cac gcc ccc 528Leu Tyr Gly Pro Asp Asn Gln Phe Glu Ala Val Ile Arg His Ala Pro 165 170 175 ttt agc gcc gac ttc aag agg gat ggc caa acc cac gtt caa ttg aac 576Phe Ser Ala Asp Phe Lys Arg Asp Gly Gln Thr His Val Gln Leu Asn 180 185 190 aac aag ggc tac ctt aac atg gag cat tgg cgc cct aag gta gag gtc 624Asn Lys Gly Tyr Leu Asn Met Glu His Trp Arg Pro Lys Val Glu Val 195 200 205 gaa ggc gag ggc gag cag caa acc cag gaa gat gaa agc act tgg tgg 672Glu Gly Glu Gly Glu Gln Gln Thr Gln Glu Asp Glu Ser Thr Trp Trp 210 215 220 gat gag agc ttt ggt gga aac acg gac acc aag ccc agg ggt ccc gag 720Asp Glu Ser Phe Gly Gly Asn Thr Asp Thr Lys Pro Arg Gly Pro Glu 225 230 235 240 agt gtg gga ttg gat atc acc ttc cct ggc tac aag cat gtt ttt gga 768Ser Val Gly Leu Asp Ile Thr Phe Pro Gly Tyr Lys His Val Phe Gly 245 250 255 att cct gag cat gct gac tct ctc tcc tta aag gaa act cga ggt ggt 816Ile Pro Glu His Ala Asp Ser Leu Ser Leu Lys Glu Thr Arg Gly Gly 260 265 270 gaa ggg aat cac gaa gag ccc tac cgc atg tac aat gcg gat gta ttt 864Glu Gly Asn His Glu Glu Pro Tyr Arg Met Tyr Asn Ala Asp Val Phe 275 280 285 gag tac gag ctg agc agt ccc atg acc ttg tat ggt gct att cca ttc 912Glu Tyr Glu Leu Ser Ser Pro Met Thr Leu Tyr Gly Ala Ile Pro Phe 290 295 300 atg cag gca cat cgc aag gac tcc acc gtc ggt gtc ttc tgg ctg aat 960Met Gln Ala His Arg Lys Asp Ser Thr Val Gly Val Phe Trp Leu Asn 305 310 315 320 gct gca gag acc tgg gtg gac att gtc aag tct acc tca tct cct aac 1008Ala Ala Glu Thr Trp Val Asp Ile Val Lys Ser Thr Ser Ser Pro Asn 325 330 335 cct ctt gct ctc ggc gtg ggc gcc acc act gac acc cag agt cat tgg 1056Pro Leu Ala Leu Gly Val Gly Ala Thr Thr Asp Thr Gln Ser His Trp 340 345 350 ttt tcg gag tcc ggc cag ctc gac gtg ttc gtt ttc ctt ggt cct acc 1104Phe Ser Glu Ser Gly Gln Leu Asp Val Phe Val Phe Leu Gly Pro Thr 355 360 365 cca cag gaa atc agc aag acc tat ggt gaa ctc acc ggc tac act cag 1152Pro Gln Glu Ile Ser Lys Thr Tyr Gly Glu Leu Thr Gly Tyr Thr Gln 370 375 380 ttg cct caa cat ttt gcc att gct tat cac cag tgc cgc tgg aac tac 1200Leu Pro Gln His Phe Ala Ile Ala Tyr His Gln Cys Arg Trp Asn Tyr 385 390 395 400 atc act gat gag gat gtc aag gag gtc gat cgc aac ttt gac aag tac 1248Ile Thr Asp Glu Asp Val Lys Glu Val Asp Arg Asn Phe Asp Lys Tyr 405 410 415 cag atc ccc tac gat gtc atc tgg ctg gac atc gaa tat acc gat gac 1296Gln Ile Pro Tyr Asp Val Ile Trp Leu Asp Ile Glu Tyr Thr Asp Asp 420 425 430 aga aag tat ttc acc tgg gat cca ctc agt ttc ccc gat ccg atc agc 1344Arg Lys Tyr Phe Thr Trp Asp Pro Leu Ser Phe Pro Asp Pro Ile Ser 435 440 445 atg gag gag cag ctc gat gag tcg gag cgc aaa ctc gtc gtt atc att 1392Met Glu Glu Gln Leu Asp Glu Ser Glu Arg Lys Leu Val Val Ile Ile 450 455 460 gac ccg cac atc aag aac cag gac aag tac agc atc gtc caa gaa atg 1440Asp Pro His Ile Lys Asn Gln Asp Lys Tyr Ser Ile Val Gln Glu Met 465 470 475 480 aag agc aaa gac ttg gcc act aag aac aag gac ggt gag atc tac gac 1488Lys Ser Lys Asp Leu Ala Thr Lys Asn Lys Asp Gly Glu Ile Tyr Asp 485 490 495 ggg tgg tgt tgg cct ggc tct tct cac tgg atc gat acc ttc aac ccc 1536Gly Trp Cys Trp Pro Gly Ser Ser His Trp Ile Asp Thr Phe Asn Pro 500 505 510 gcc gcc atc aaa tgg tgg gtc agc tta ttc aag ttt gac aag ttc aag 1584Ala Ala Ile Lys Trp Trp Val Ser Leu Phe Lys Phe Asp Lys Phe Lys 515 520 525 ggg acg ctg tcc aat gtc ttc att tgg aac gac atg aac gag ccc tcg 1632Gly Thr Leu Ser Asn Val Phe Ile Trp Asn Asp Met Asn Glu Pro Ser 530 535 540 gtt ttc aac ggt ccc gaa acc acg atg ccc aag gat aac ctt cat cat 1680Val Phe Asn Gly Pro Glu Thr Thr Met Pro Lys Asp Asn Leu His His 545 550 555 560 ggc aac tgg gag cac cgt gac atc cat aac gtt cat gga atc acc ctg 1728Gly Asn Trp Glu His Arg Asp Ile His Asn Val His Gly Ile Thr Leu 565 570 575 gtc aat gcc acc tac gat gcc ctt cta gag cgc aag aag ggc gag atc 1776Val Asn Ala Thr Tyr Asp Ala Leu Leu Glu Arg Lys Lys Gly Glu Ile 580 585 590 cgt cgg cct ttc att ctg aca cgg tca tat tat gct ggt gct caa cgg 1824Arg Arg Pro Phe Ile Leu Thr Arg Ser Tyr Tyr Ala Gly Ala Gln Arg 595 600 605 atg tct gct atg tgg acg ggt gat aac cag gct act tgg gaa cac ttg 1872Met Ser Ala Met Trp Thr Gly Asp Asn Gln Ala Thr Trp Glu His Leu 610 615 620 gcc gct tcc atc cct atg gtt ctg aac aac ggc att gcg ggc ttc ccc 1920Ala Ala Ser Ile Pro Met Val Leu Asn Asn Gly Ile Ala Gly Phe Pro 625 630 635 640 ttt gcc ggt gct gac gtg ggc ggt ttc ttc cag aac cct agc aag gag 1968Phe Ala Gly Ala Asp Val Gly Gly Phe Phe Gln Asn Pro Ser Lys Glu 645 650 655 ctc ttg acc aga tgg tac caa gct ggt att tgg tac ccc ttc ttc cgg 2016Leu Leu Thr Arg Trp Tyr Gln Ala Gly Ile Trp Tyr Pro Phe Phe Arg 660 665 670 gcc cac gcg cat att gac acg cgc cgg aga gag ccg tat ctg att gcc 2064Ala His Ala His Ile Asp Thr Arg Arg Arg Glu Pro Tyr Leu Ile Ala 675 680 685 gag cca cac cgg tct atc atc tcc cag gct atc cgc ctg agg tat cag 2112Glu Pro His Arg Ser Ile Ile Ser Gln Ala Ile Arg Leu Arg Tyr Gln 690 695 700 ctt ctc ccc gcc tgg tac act gcc ttc cac gaa gct tcc gtg aac gga 2160Leu Leu Pro Ala Trp Tyr Thr Ala Phe His Glu Ala Ser Val Asn Gly 705 710 715 720 atg ccg atc gtg agg ccg cag tac tac gct cac cct tgg gat gag gct 2208Met Pro Ile Val Arg Pro Gln Tyr Tyr Ala His Pro Trp Asp Glu Ala 725 730 735 ggc ttt gcc att gac gac cag ctt tat ctc ggc tcc acc ggt ctt ctt 2256Gly Phe Ala Ile Asp Asp Gln Leu Tyr Leu Gly Ser Thr Gly Leu Leu 740 745 750 gct aag cct gtt gtc tcc gag gag gcc acc acg gcc gac att tac ctt 2304Ala Lys Pro Val Val Ser Glu Glu Ala Thr Thr Ala Asp Ile Tyr Leu 755 760 765 gct gac gac gaa aag tac tat gac tac ttt gac tac acc gtc tac cag 2352Ala Asp Asp Glu Lys Tyr Tyr Asp Tyr Phe Asp Tyr Thr Val Tyr Gln 770 775 780 gga gcc gga aag cgg cat acg gtg cct gct cct atg gag act gtg cca 2400Gly Ala Gly Lys Arg His Thr Val Pro Ala Pro Met Glu Thr Val Pro 785 790 795 800 ttg ctg atg cag ggt ggc cat gta atc ccc cgc aag gac cgt cct cgc 2448Leu Leu Met Gln Gly Gly His Val Ile Pro Arg Lys Asp Arg Pro Arg 805 810 815 cgc agt agc gcc ttg atg aga tgg gat ccg tac act ctt gtt gtg gtc 2496Arg Ser Ser Ala Leu Met Arg Trp Asp Pro Tyr Thr Leu Val Val Val 820 825 830 ttg gat aag aac ggt caa gcc gat ggc tct ctc tac gtg gat gac ggt 2544Leu Asp Lys Asn Gly Gln Ala Asp Gly Ser Leu Tyr Val Asp Asp Gly 835 840 845 gag acg ttc gac tat gag cgt gga gct tat atc cac cgc cgt ttc cgc 2592Glu Thr Phe Asp Tyr Glu Arg Gly Ala Tyr Ile His Arg Arg Phe Arg 850 855 860 ttc cag gag tct gcc ctg gtc tcg gag gat gtt ggc acc aag ggt cct 2640Phe Gln Glu Ser Ala Leu Val Ser Glu Asp Val Gly Thr Lys Gly Pro 865 870 875 880 aag acg gcc gag tac ttg aag acc atg gcc aac gtt cgt gtt gag cgg 2688Lys Thr Ala Glu Tyr Leu Lys Thr Met Ala Asn Val Arg Val Glu Arg 885 890 895 gtg gtg gta gtt gat cct cct aag gaa tgg cag ggt aag acc agt gtg 2736Val Val Val Val Asp Pro Pro Lys Glu Trp Gln Gly Lys Thr Ser Val 900 905 910 act gtc att gag gat gga gct tcg gcg gct tcg aca gcc tct atg cag 2784Thr Val Ile Glu Asp Gly Ala Ser Ala Ala Ser Thr Ala Ser Met Gln 915 920 925 tac cac agc cag ccc gat ggc aag

gcc gca tat gcg gtg gtg aag aac 2832Tyr His Ser Gln Pro Asp Gly Lys Ala Ala Tyr Ala Val Val Lys Asn 930 935 940 ccc aat gtc ggc att gga aag aca tgg cgg att gag ttt tag 2874Pro Asn Val Gly Ile Gly Lys Thr Trp Arg Ile Glu Phe 945 950 955 30957PRTAspergillus niger 30Met Ser Asn Arg Trp Thr Leu Leu Leu Ser Leu Val Ile Leu Leu Gly 1 5 10 15 Cys Leu Val Ile Pro Gly Val Thr Val Lys His Glu Asn Phe Lys Thr 20 25 30 Cys Ser Gln Ser Gly Phe Cys Lys Arg Asn Arg Ala Phe Ala Asp Asp 35 40 45 Ala Ala Ala Gln Gly Ser Ser Trp Ala Ser Pro Tyr Glu Leu Asp Ser 50 55 60 Ser Ser Ile Gln Phe Lys Asp Gly Gln Leu His Gly Thr Ile Leu Lys 65 70 75 80 Ser Val Ser Pro Asn Glu Lys Val Lys Leu Pro Leu Val Val Ser Phe 85 90 95 Leu Glu Ser Gly Ala Ala Arg Val Val Val Asp Glu Glu Lys Arg Met 100 105 110 Asn Gly Asp Ile Gln Leu Arg His Asp Ser Lys Ala Arg Lys Glu Arg 115 120 125 Tyr Asn Glu Ala Glu Lys Trp Val Leu Val Gly Gly Leu Glu Leu Ser 130 135 140 Lys Thr Ala Thr Leu Arg Pro Glu Thr Glu Ser Gly Phe Thr Arg Val 145 150 155 160 Leu Tyr Gly Pro Asp Asn Gln Phe Glu Ala Val Ile Arg His Ala Pro 165 170 175 Phe Ser Ala Asp Phe Lys Arg Asp Gly Gln Thr His Val Gln Leu Asn 180 185 190 Asn Lys Gly Tyr Leu Asn Met Glu His Trp Arg Pro Lys Val Glu Val 195 200 205 Glu Gly Glu Gly Glu Gln Gln Thr Gln Glu Asp Glu Ser Thr Trp Trp 210 215 220 Asp Glu Ser Phe Gly Gly Asn Thr Asp Thr Lys Pro Arg Gly Pro Glu 225 230 235 240 Ser Val Gly Leu Asp Ile Thr Phe Pro Gly Tyr Lys His Val Phe Gly 245 250 255 Ile Pro Glu His Ala Asp Ser Leu Ser Leu Lys Glu Thr Arg Gly Gly 260 265 270 Glu Gly Asn His Glu Glu Pro Tyr Arg Met Tyr Asn Ala Asp Val Phe 275 280 285 Glu Tyr Glu Leu Ser Ser Pro Met Thr Leu Tyr Gly Ala Ile Pro Phe 290 295 300 Met Gln Ala His Arg Lys Asp Ser Thr Val Gly Val Phe Trp Leu Asn 305 310 315 320 Ala Ala Glu Thr Trp Val Asp Ile Val Lys Ser Thr Ser Ser Pro Asn 325 330 335 Pro Leu Ala Leu Gly Val Gly Ala Thr Thr Asp Thr Gln Ser His Trp 340 345 350 Phe Ser Glu Ser Gly Gln Leu Asp Val Phe Val Phe Leu Gly Pro Thr 355 360 365 Pro Gln Glu Ile Ser Lys Thr Tyr Gly Glu Leu Thr Gly Tyr Thr Gln 370 375 380 Leu Pro Gln His Phe Ala Ile Ala Tyr His Gln Cys Arg Trp Asn Tyr 385 390 395 400 Ile Thr Asp Glu Asp Val Lys Glu Val Asp Arg Asn Phe Asp Lys Tyr 405 410 415 Gln Ile Pro Tyr Asp Val Ile Trp Leu Asp Ile Glu Tyr Thr Asp Asp 420 425 430 Arg Lys Tyr Phe Thr Trp Asp Pro Leu Ser Phe Pro Asp Pro Ile Ser 435 440 445 Met Glu Glu Gln Leu Asp Glu Ser Glu Arg Lys Leu Val Val Ile Ile 450 455 460 Asp Pro His Ile Lys Asn Gln Asp Lys Tyr Ser Ile Val Gln Glu Met 465 470 475 480 Lys Ser Lys Asp Leu Ala Thr Lys Asn Lys Asp Gly Glu Ile Tyr Asp 485 490 495 Gly Trp Cys Trp Pro Gly Ser Ser His Trp Ile Asp Thr Phe Asn Pro 500 505 510 Ala Ala Ile Lys Trp Trp Val Ser Leu Phe Lys Phe Asp Lys Phe Lys 515 520 525 Gly Thr Leu Ser Asn Val Phe Ile Trp Asn Asp Met Asn Glu Pro Ser 530 535 540 Val Phe Asn Gly Pro Glu Thr Thr Met Pro Lys Asp Asn Leu His His 545 550 555 560 Gly Asn Trp Glu His Arg Asp Ile His Asn Val His Gly Ile Thr Leu 565 570 575 Val Asn Ala Thr Tyr Asp Ala Leu Leu Glu Arg Lys Lys Gly Glu Ile 580 585 590 Arg Arg Pro Phe Ile Leu Thr Arg Ser Tyr Tyr Ala Gly Ala Gln Arg 595 600 605 Met Ser Ala Met Trp Thr Gly Asp Asn Gln Ala Thr Trp Glu His Leu 610 615 620 Ala Ala Ser Ile Pro Met Val Leu Asn Asn Gly Ile Ala Gly Phe Pro 625 630 635 640 Phe Ala Gly Ala Asp Val Gly Gly Phe Phe Gln Asn Pro Ser Lys Glu 645 650 655 Leu Leu Thr Arg Trp Tyr Gln Ala Gly Ile Trp Tyr Pro Phe Phe Arg 660 665 670 Ala His Ala His Ile Asp Thr Arg Arg Arg Glu Pro Tyr Leu Ile Ala 675 680 685 Glu Pro His Arg Ser Ile Ile Ser Gln Ala Ile Arg Leu Arg Tyr Gln 690 695 700 Leu Leu Pro Ala Trp Tyr Thr Ala Phe His Glu Ala Ser Val Asn Gly 705 710 715 720 Met Pro Ile Val Arg Pro Gln Tyr Tyr Ala His Pro Trp Asp Glu Ala 725 730 735 Gly Phe Ala Ile Asp Asp Gln Leu Tyr Leu Gly Ser Thr Gly Leu Leu 740 745 750 Ala Lys Pro Val Val Ser Glu Glu Ala Thr Thr Ala Asp Ile Tyr Leu 755 760 765 Ala Asp Asp Glu Lys Tyr Tyr Asp Tyr Phe Asp Tyr Thr Val Tyr Gln 770 775 780 Gly Ala Gly Lys Arg His Thr Val Pro Ala Pro Met Glu Thr Val Pro 785 790 795 800 Leu Leu Met Gln Gly Gly His Val Ile Pro Arg Lys Asp Arg Pro Arg 805 810 815 Arg Ser Ser Ala Leu Met Arg Trp Asp Pro Tyr Thr Leu Val Val Val 820 825 830 Leu Asp Lys Asn Gly Gln Ala Asp Gly Ser Leu Tyr Val Asp Asp Gly 835 840 845 Glu Thr Phe Asp Tyr Glu Arg Gly Ala Tyr Ile His Arg Arg Phe Arg 850 855 860 Phe Gln Glu Ser Ala Leu Val Ser Glu Asp Val Gly Thr Lys Gly Pro 865 870 875 880 Lys Thr Ala Glu Tyr Leu Lys Thr Met Ala Asn Val Arg Val Glu Arg 885 890 895 Val Val Val Val Asp Pro Pro Lys Glu Trp Gln Gly Lys Thr Ser Val 900 905 910 Thr Val Ile Glu Asp Gly Ala Ser Ala Ala Ser Thr Ala Ser Met Gln 915 920 925 Tyr His Ser Gln Pro Asp Gly Lys Ala Ala Tyr Ala Val Val Lys Asn 930 935 940 Pro Asn Val Gly Ile Gly Lys Thr Trp Arg Ile Glu Phe 945 950 955 312303DNAAspergillus niger 31agcggcaggc cgataaggag ctgagtcagc gagccccaga atgggcgggc gattatcacc 60agctggccag aagctctctc cgcattcctg atggtggatt gattgatgaa ttgattgctt 120ttttgtcttg ctcttgttag tttgttctct agtgccccta cacagcattg gtcagggagc 180tcaatgctgc cactgccacc atgatacttc ctcagggatc gctcttcttg gtgagcatag 240ctgcttgctc gaccgtcgtg gctgcggcgg gtgatgcctc ctctcgtccc cggggtgtag 300gtcccgaatg taagtgccac ttatatacct acttaggctg aggcggtgct acacgcaacc 360attctattcc aacttgcatc ctcgccataa tcttgttttt ttgtcggcat tggtcatgct 420aactggtttc gtctaatggt tggcagtcgc caagttctac aaggatacca ccaccttcac 480gtgcatctcc cacccagcca tccagatccc cttctccgcc gtgaacgatg attactgtga 540ctgtccggat ggcagtgatg agcctggcac atctgcctgt gccttcctgt ctcgcaactc 600cgccctaaca ccgggtgagc gccccggcag cgacgatctc gagctgacat ccgccctgcc 660gggtttctac tgcaagaaca agggccacaa gcccggctac gtccccttcc agcgggtcaa 720tgacggcatc tgtgactatg agctctgctg cgacggcagt gacgagtggg cccgccctgg 780cggcaccaag tgtgaagaca agtgcaagga gatcggcaag gaatggcgga agaaggagga 840gaagagacag aagtccatga ctgcggcttt gaagaagaag aaggatctgc ttgtggaggc 900tggtagacag cagaaggagg tcgaggacaa tatcaagcgt ctggaagttg aaattcaggc 960ccaggagctg aaggtcaatg atcttcaggc ggagctggag gaggtggagc agcaggaggc 1020gagcaaggtc gtgaagggca agacggcggg caaggttaat gtgcttgctg ggttggctaa 1080gagccgggtt gaggagcttc gaaacgccct gatggacgtc cgcaaggagc gtgatgatac 1140ccgtgcccgt gtgaaggagc tcgaagagat tctgtctaag ttcaaggtgg aatacaaccc 1200taacttcaac gatgagggcg ttaagcgcgc tgtgcgcagc tgggaagact acgccgccaa 1260gggcaccctt gagggcgccg tgaacaacgc tcaggaccgt gatttggatg aaattgctaa 1320gcccgatgat gagaaggcgg gcatcaactg ggaacagtgg gagaatgaag aggatgggtg 1380tgaggctggt cttggtatgt aaatcatttc agagtgaggg ttatcgatgt tcccgcgcta 1440acatcagatt tagtctacca gctggcagcc taccttccgc cttctttggt cgagtttatc 1500gaaggcaagg tgctcttcgt cagaggtctc ttggaagata acggaattct acccaaggcg 1560gccgagactt ctacgtccga atccaaggtt gtgtcagaag cccgagaagc cgtgaagtca 1620gcagagaagg agcttggaga caagcagaag cagctgaagg atcacaagtc cgatcttgag 1680acggactatg gtgtcggatc catcttccgt gccctcaagg gcgtttgcat ctccaaggac 1740tcgggtgagt acacgtatga gcactgcttc ctggaccaga caaaacagat tccaaagaag 1800ggcggcggat ccacacgcat gggcaagtac accggcattg ggtcggtcag tgttgatgtg 1860ctcaacgagg cgggcgagat tgtccccgaa gacagggtca ctcttcagta cgccaacgga 1920caaggctgct ggaatggacc ggcccgctcg acgacggtca tcctgacatg cggcgaagag 1980gatgcgatcc tgaaggtggc cgaagacgag aagtgcgtgt actcgatgca tgtcacgtcg 2040ccggccgtgt gtcccggagg cgatgagggc gcaactgccc cgaaccgcaa ggatgagctg 2100tgagcagtga tgggaccata tttagggtta tatcagagcg ctacaagtct ataggttttg 2160cttatttgaa ttgcatacac agcatctgtt gttctgacag caatcatgag ctactgacct 2220aatggcctta gtattgacta ggcagctgcc aactagcaga actgctgatt agcgggtggc 2280atcggagtag cttctgttgc tta 2303321707DNAAspergillus nigerCDS(1)..(1707) 32atg ata ctt cct cag gga tcg ctc ttc ttg gtg agc ata gct gct tgc 48Met Ile Leu Pro Gln Gly Ser Leu Phe Leu Val Ser Ile Ala Ala Cys 1 5 10 15 tcg acc gtc gtg gct gcg gcg ggt gat gcc tcc tct cgt ccc cgg ggt 96Ser Thr Val Val Ala Ala Ala Gly Asp Ala Ser Ser Arg Pro Arg Gly 20 25 30 gta ggt ccc gaa ttc gcc aag ttc tac aag gat acc acc acc ttc acg 144Val Gly Pro Glu Phe Ala Lys Phe Tyr Lys Asp Thr Thr Thr Phe Thr 35 40 45 tgc atc tcc cac cca gcc atc cag atc ccc ttc tcc gcc gtg aac gat 192Cys Ile Ser His Pro Ala Ile Gln Ile Pro Phe Ser Ala Val Asn Asp 50 55 60 gat tac tgt gac tgt ccg gat ggc agt gat gag cct ggc aca tct gcc 240Asp Tyr Cys Asp Cys Pro Asp Gly Ser Asp Glu Pro Gly Thr Ser Ala 65 70 75 80 tgt gcc ttc ctg tct cgc aac tcc gcc cta aca ccg ggt gag cgc ccc 288Cys Ala Phe Leu Ser Arg Asn Ser Ala Leu Thr Pro Gly Glu Arg Pro 85 90 95 ggc agc gac gat ctc gag ctg aca tcc gcc ctg ccg ggt ttc tac tgc 336Gly Ser Asp Asp Leu Glu Leu Thr Ser Ala Leu Pro Gly Phe Tyr Cys 100 105 110 aag aac aag ggc cac aag ccc ggc tac gtc ccc ttc cag cgg gtc aat 384Lys Asn Lys Gly His Lys Pro Gly Tyr Val Pro Phe Gln Arg Val Asn 115 120 125 gac ggc atc tgt gac tat gag ctc tgc tgc gac ggc agt gac gag tgg 432Asp Gly Ile Cys Asp Tyr Glu Leu Cys Cys Asp Gly Ser Asp Glu Trp 130 135 140 gcc cgc cct ggc ggc acc aag tgt gaa gac aag tgc aag gag atc ggc 480Ala Arg Pro Gly Gly Thr Lys Cys Glu Asp Lys Cys Lys Glu Ile Gly 145 150 155 160 aag gaa tgg cgg aag aag gag gag aag aga cag aag tcc atg act gcg 528Lys Glu Trp Arg Lys Lys Glu Glu Lys Arg Gln Lys Ser Met Thr Ala 165 170 175 gct ttg aag aag aag aag gat ctg ctt gtg gag gct ggt aga cag cag 576Ala Leu Lys Lys Lys Lys Asp Leu Leu Val Glu Ala Gly Arg Gln Gln 180 185 190 aag gag gtc gag gac aat atc aag cgt ctg gaa gtt gaa att cag gcc 624Lys Glu Val Glu Asp Asn Ile Lys Arg Leu Glu Val Glu Ile Gln Ala 195 200 205 cag gag ctg aag gtc aat gat ctt cag gcg gag ctg gag gag gtg gag 672Gln Glu Leu Lys Val Asn Asp Leu Gln Ala Glu Leu Glu Glu Val Glu 210 215 220 cag cag gag gcg agc aag gtc gtg aag ggc aag acg gcg ggc aag gtt 720Gln Gln Glu Ala Ser Lys Val Val Lys Gly Lys Thr Ala Gly Lys Val 225 230 235 240 aat gtg ctt gct ggg ttg gct aag agc cgg gtt gag gag ctt cga aac 768Asn Val Leu Ala Gly Leu Ala Lys Ser Arg Val Glu Glu Leu Arg Asn 245 250 255 gcc ctg atg gac gtc cgc aag gag cgt gat gat acc cgt gcc cgt gtg 816Ala Leu Met Asp Val Arg Lys Glu Arg Asp Asp Thr Arg Ala Arg Val 260 265 270 aag gag ctc gaa gag att ctg tct aag ttc aag gtg gaa tac aac cct 864Lys Glu Leu Glu Glu Ile Leu Ser Lys Phe Lys Val Glu Tyr Asn Pro 275 280 285 aac ttc aac gat gag ggc gtt aag cgc gct gtg cgc agc tgg gaa gac 912Asn Phe Asn Asp Glu Gly Val Lys Arg Ala Val Arg Ser Trp Glu Asp 290 295 300 tac gcc gcc aag ggc acc ctt gag ggc gcc gtg aac aac gct cag gac 960Tyr Ala Ala Lys Gly Thr Leu Glu Gly Ala Val Asn Asn Ala Gln Asp 305 310 315 320 cgt gat ttg gat gaa att gct aag ccc gat gat gag aag gcg ggc atc 1008Arg Asp Leu Asp Glu Ile Ala Lys Pro Asp Asp Glu Lys Ala Gly Ile 325 330 335 aac tgg gaa cag tgg gag aat gaa gag gat ggg tgt gag gct ggt ctt 1056Asn Trp Glu Gln Trp Glu Asn Glu Glu Asp Gly Cys Glu Ala Gly Leu 340 345 350 gtc tac cag ctg gca gcc tac ctt ccg cct tct ttg gtc gag ttt atc 1104Val Tyr Gln Leu Ala Ala Tyr Leu Pro Pro Ser Leu Val Glu Phe Ile 355 360 365 gaa ggc aag gtg ctc ttc gtc aga ggt ctc ttg gaa gat aac gga att 1152Glu Gly Lys Val Leu Phe Val Arg Gly Leu Leu Glu Asp Asn Gly Ile 370 375 380 cta ccc aag gcg gcc gag act tct acg tcc gaa tcc aag gtt gtg tca 1200Leu Pro Lys Ala Ala Glu Thr Ser Thr Ser Glu Ser Lys Val Val Ser 385 390 395 400 gaa gcc cga gaa gcc gtg aag tca gca gag aag gag ctt gga gac aag 1248Glu Ala Arg Glu Ala Val Lys Ser Ala Glu Lys Glu Leu Gly Asp Lys 405 410 415 cag aag cag ctg aag gat cac aag tcc gat ctt gag acg gac tat ggt 1296Gln Lys Gln Leu Lys Asp His Lys Ser Asp Leu Glu Thr Asp Tyr Gly 420 425 430 gtc gga tcc atc ttc cgt gcc ctc aag ggc gtt tgc atc tcc aag gac 1344Val Gly Ser Ile Phe Arg Ala Leu Lys Gly Val Cys Ile Ser Lys Asp 435 440 445 tcg ggt gag tac acg tat gag cac tgc ttc ctg gac cag aca aaa cag 1392Ser Gly Glu Tyr Thr Tyr Glu His Cys Phe Leu Asp Gln Thr Lys Gln 450 455 460 att cca aag aag ggc ggc gga tcc aca cgc atg ggc aag tac acc ggc 1440Ile Pro Lys Lys Gly Gly Gly Ser Thr Arg Met Gly Lys Tyr Thr Gly 465 470 475 480 att ggg tcg gtc agt gtt gat gtg ctc aac gag gcg ggc gag att gtc 1488Ile Gly Ser Val Ser Val Asp Val Leu Asn Glu Ala Gly Glu Ile Val 485 490 495 ccc gaa gac agg gtc act ctt cag tac gcc aac gga caa ggc tgc tgg 1536Pro Glu Asp Arg Val Thr Leu Gln Tyr Ala Asn Gly Gln Gly Cys Trp 500 505 510 aat gga ccg gcc cgc tcg acg acg gtc atc ctg aca tgc ggc gaa gag 1584Asn Gly Pro Ala Arg Ser Thr Thr Val Ile Leu Thr Cys Gly Glu Glu 515 520 525 gat gcg atc ctg aag gtg gcc gaa gac gag aag tgc gtg tac tcg atg 1632Asp Ala Ile Leu Lys Val Ala Glu Asp Glu Lys Cys Val Tyr Ser Met 530 535 540 cat gtc acg tcg ccg gcc gtg tgt ccc gga ggc gat gag ggc gca act 1680His Val Thr Ser Pro Ala Val Cys Pro Gly Gly Asp Glu Gly Ala Thr 545 550 555 560 gcc ccg aac cgc aag gat gag ctg tga 1707Ala Pro Asn Arg Lys Asp Glu Leu 565 33568PRTAspergillus niger 33Met Ile Leu Pro Gln Gly Ser Leu Phe Leu Val Ser Ile Ala Ala Cys 1 5 10 15 Ser Thr Val Val Ala Ala Ala Gly Asp Ala Ser Ser Arg Pro Arg Gly 20 25

30 Val Gly Pro Glu Phe Ala Lys Phe Tyr Lys Asp Thr Thr Thr Phe Thr 35 40 45 Cys Ile Ser His Pro Ala Ile Gln Ile Pro Phe Ser Ala Val Asn Asp 50 55 60 Asp Tyr Cys Asp Cys Pro Asp Gly Ser Asp Glu Pro Gly Thr Ser Ala 65 70 75 80 Cys Ala Phe Leu Ser Arg Asn Ser Ala Leu Thr Pro Gly Glu Arg Pro 85 90 95 Gly Ser Asp Asp Leu Glu Leu Thr Ser Ala Leu Pro Gly Phe Tyr Cys 100 105 110 Lys Asn Lys Gly His Lys Pro Gly Tyr Val Pro Phe Gln Arg Val Asn 115 120 125 Asp Gly Ile Cys Asp Tyr Glu Leu Cys Cys Asp Gly Ser Asp Glu Trp 130 135 140 Ala Arg Pro Gly Gly Thr Lys Cys Glu Asp Lys Cys Lys Glu Ile Gly 145 150 155 160 Lys Glu Trp Arg Lys Lys Glu Glu Lys Arg Gln Lys Ser Met Thr Ala 165 170 175 Ala Leu Lys Lys Lys Lys Asp Leu Leu Val Glu Ala Gly Arg Gln Gln 180 185 190 Lys Glu Val Glu Asp Asn Ile Lys Arg Leu Glu Val Glu Ile Gln Ala 195 200 205 Gln Glu Leu Lys Val Asn Asp Leu Gln Ala Glu Leu Glu Glu Val Glu 210 215 220 Gln Gln Glu Ala Ser Lys Val Val Lys Gly Lys Thr Ala Gly Lys Val 225 230 235 240 Asn Val Leu Ala Gly Leu Ala Lys Ser Arg Val Glu Glu Leu Arg Asn 245 250 255 Ala Leu Met Asp Val Arg Lys Glu Arg Asp Asp Thr Arg Ala Arg Val 260 265 270 Lys Glu Leu Glu Glu Ile Leu Ser Lys Phe Lys Val Glu Tyr Asn Pro 275 280 285 Asn Phe Asn Asp Glu Gly Val Lys Arg Ala Val Arg Ser Trp Glu Asp 290 295 300 Tyr Ala Ala Lys Gly Thr Leu Glu Gly Ala Val Asn Asn Ala Gln Asp 305 310 315 320 Arg Asp Leu Asp Glu Ile Ala Lys Pro Asp Asp Glu Lys Ala Gly Ile 325 330 335 Asn Trp Glu Gln Trp Glu Asn Glu Glu Asp Gly Cys Glu Ala Gly Leu 340 345 350 Val Tyr Gln Leu Ala Ala Tyr Leu Pro Pro Ser Leu Val Glu Phe Ile 355 360 365 Glu Gly Lys Val Leu Phe Val Arg Gly Leu Leu Glu Asp Asn Gly Ile 370 375 380 Leu Pro Lys Ala Ala Glu Thr Ser Thr Ser Glu Ser Lys Val Val Ser 385 390 395 400 Glu Ala Arg Glu Ala Val Lys Ser Ala Glu Lys Glu Leu Gly Asp Lys 405 410 415 Gln Lys Gln Leu Lys Asp His Lys Ser Asp Leu Glu Thr Asp Tyr Gly 420 425 430 Val Gly Ser Ile Phe Arg Ala Leu Lys Gly Val Cys Ile Ser Lys Asp 435 440 445 Ser Gly Glu Tyr Thr Tyr Glu His Cys Phe Leu Asp Gln Thr Lys Gln 450 455 460 Ile Pro Lys Lys Gly Gly Gly Ser Thr Arg Met Gly Lys Tyr Thr Gly 465 470 475 480 Ile Gly Ser Val Ser Val Asp Val Leu Asn Glu Ala Gly Glu Ile Val 485 490 495 Pro Glu Asp Arg Val Thr Leu Gln Tyr Ala Asn Gly Gln Gly Cys Trp 500 505 510 Asn Gly Pro Ala Arg Ser Thr Thr Val Ile Leu Thr Cys Gly Glu Glu 515 520 525 Asp Ala Ile Leu Lys Val Ala Glu Asp Glu Lys Cys Val Tyr Ser Met 530 535 540 His Val Thr Ser Pro Ala Val Cys Pro Gly Gly Asp Glu Gly Ala Thr 545 550 555 560 Ala Pro Asn Arg Lys Asp Glu Leu 565 342343DNAAspergillus niger 34gcctgaccac gatcgaccgt tgacaatcca ggcttccctt ccgtcgctga tcttcatgtg 60aaggaccctg ttttcttgct tacctaccat agaccttcat tgcatagctt cactctatac 120tctcttggtt tctgtcatac actccaattc cttcctccgt ccttgtccac tcgtttccgc 180cggtctccgg ctattccaag atgcgctccg cagccaaatt tttctatttg gcggtatttg 240ccctgtcgag gctttccaat gccgagactg ggctgcacca taaccaggac aagtgtgcgg 300taagtcgata caacgctcgg gcatacgctt ggactgcatg ttgtagaccg gacttggctg 360attcattctc atctctacca gattgacccc actgccatgg tgtccgacgc ttgtgtctcc 420tacgccacta tcgatcatct gaacgatcaa gtctacaccc tcctccaatc cattacgcaa 480gataccgatt tcttctcgta ctaccgtctt aatctcttca acaaagtctg tccattctgg 540tccgatgcga atagtatgtg cgggaacatt gcatgctccg tcaacacaat cgaatctgaa 600gacgacattc cgttaacatg gcgcgcggag gagctcagta aactcgaggg acccaaagca 660ggccatccgg gccgcaatca acgaaaggag cgacctctta atcgaccgct ccaaggaatg 720ctaggcgaaa atgttggaga gagctgtgtg gtggagtatg acgatgaatg tgatgaacgg 780gactactgtg ttcccgaaga tgagggtgct agcggcaagg gagactatgt cagtctcgtt 840gataatccag aacggttcac agggtatgcc ggtatgggcg cccatcaggt ttgggatgca 900atctatcggg agaattgctt cctcaaaccg gtgcccgagc tatcacccgt tactcctcag 960ctgggtggtc ttcaagctgt caacgatttc cgtcatgtgc ttcagcagga gttgaagcgc 1020cctgacctgc ttccattgga caatgaatgc cttgagaagc gagtgttcca tcgtctcatc 1080agcggaatgc atgcgtctat ctcgacccac ctttgctggg actacctaaa ccagacgacg 1140ggacaatggc atcctaacct tcaatgcttc aaagatcgtc tccacgatca ccccgagcgc 1200atctcgaacc tgtacttcaa ctacgcgctg gtctcgcgcg ccgtggcgaa gctgcagaaa 1260cacctacaca actacaacta ctgcgtcggt gatccggtcc aggatgccat gactagggag 1320aaggtctcca agttgacctc gaccttggct gaccgccctc aaattttcga cgagaacgtc 1380atgttccagg atcccagctc cgctggcctg aaggaagact tccgcaaccg attccgcaac 1440gtcagtcgcc tgatggactg cgtcgggtgc gacaaatgcc gcctctgggg caagctccag 1500gtcaacggat atggcaccgc tctgaaagtg ctgttcgagt acgacgagac taagaacggc 1560gagaacccgt tgctgcgccg gactgagctg gtggcactga tcaataccct tggtcgcatt 1620tctcacagca ttgccgccgt ccggagtttc caccgggcca tggatgtggg cgatggggag 1680gtcttcacca tccccgcgag cattgcgtcc aaggagcgcg gtggcaagaa gaagacccga 1740cgacttctca aagacggtgg ctcaaccttc tattatgagg atggcgatga tgacaacttt 1800gtctacatca ccgagaaact tccgtgggag aaggtccggg tacgccgcga cacggatacg 1860gtctgggatg atattaaggc cgagttttct atgatctggg acatttacgt ctatgtgctg 1920aagagctggg tcaatgcacc aaagacattg taagtgatgg catacgatga cctccacagt 1980ttcctatata ctaactgggc attgcagctt cgagatcgcc gtcctggagg ttgctcgggt 2040atggaactac tggctgggtc tgcctgtgcc gccacggtcc tggaggatcc agcttcccaa 2100gcgacccacc cctccaacac ccccgaccca tgaggagctc tagacggtag aagtgggcaa 2160ggcgacgtgg tggaagatgg cggtgactgg gattgtattg tataactagc gtcgttgagg 2220actaatcttt ttccgatctg tttgggccgg cgttgttgac gatgcctgtt gggaaatcgc 2280atctcgggag ttctgggagc attttaggcc gctgtacata tttcaagcaa tgggcctgag 2340cat 2343351803DNAAspergillus nigerCDS(1)..(1803) 35atg cgc tcc gca gcc aaa ttt ttc tat ttg gcg gta ttt gcc ctg tcg 48Met Arg Ser Ala Ala Lys Phe Phe Tyr Leu Ala Val Phe Ala Leu Ser 1 5 10 15 agg ctt tcc aat gcc gag act ggg ctg cac cat aac cag gac aag tgt 96Arg Leu Ser Asn Ala Glu Thr Gly Leu His His Asn Gln Asp Lys Cys 20 25 30 gcg att gac ccc act gcc atg gtg tcc gac gct tgt gtc tcc tac gcc 144Ala Ile Asp Pro Thr Ala Met Val Ser Asp Ala Cys Val Ser Tyr Ala 35 40 45 act atc gat cat ctg aac gat caa gtc tac acc ctc ctc caa tcc att 192Thr Ile Asp His Leu Asn Asp Gln Val Tyr Thr Leu Leu Gln Ser Ile 50 55 60 acg caa gat acc gat ttc ttc tcg tac tac cgt ctt aat ctc ttc aac 240Thr Gln Asp Thr Asp Phe Phe Ser Tyr Tyr Arg Leu Asn Leu Phe Asn 65 70 75 80 aaa gtc tgt cca ttc tgg tcc gat gcg aat agt atg tgc ggg aac att 288Lys Val Cys Pro Phe Trp Ser Asp Ala Asn Ser Met Cys Gly Asn Ile 85 90 95 gca tgc tcc gtc aac aca atc gaa tct gaa gac gac att ccg tta aca 336Ala Cys Ser Val Asn Thr Ile Glu Ser Glu Asp Asp Ile Pro Leu Thr 100 105 110 tgg cgc gcg gag gag ctc agt aaa ctc gag gga ccc aaa gca ggc cat 384Trp Arg Ala Glu Glu Leu Ser Lys Leu Glu Gly Pro Lys Ala Gly His 115 120 125 ccg ggc cgc aat caa cga aag gag cga cct ctt aat cga ccg ctc caa 432Pro Gly Arg Asn Gln Arg Lys Glu Arg Pro Leu Asn Arg Pro Leu Gln 130 135 140 gga atg cta ggc gaa aat gtt gga gag agc tgt gtg gtg gag tat gac 480Gly Met Leu Gly Glu Asn Val Gly Glu Ser Cys Val Val Glu Tyr Asp 145 150 155 160 gat gaa tgt gat gaa cgg gac tac tgt gtt ccc gaa gat gag ggt gct 528Asp Glu Cys Asp Glu Arg Asp Tyr Cys Val Pro Glu Asp Glu Gly Ala 165 170 175 agc ggc aag gga gac tat gtc agt ctc gtt gat aat cca gaa cgg ttc 576Ser Gly Lys Gly Asp Tyr Val Ser Leu Val Asp Asn Pro Glu Arg Phe 180 185 190 aca ggg tat gcc ggt atg ggc gcc cat cag gtt tgg gat gca atc tat 624Thr Gly Tyr Ala Gly Met Gly Ala His Gln Val Trp Asp Ala Ile Tyr 195 200 205 cgg gag aat tgc ttc ctc aaa ccg gtg ccc gag cta tca ccc gtt act 672Arg Glu Asn Cys Phe Leu Lys Pro Val Pro Glu Leu Ser Pro Val Thr 210 215 220 cct cag ctg ggt ggt ctt caa gct gtc aac gat ttc cgt cat gtg ctt 720Pro Gln Leu Gly Gly Leu Gln Ala Val Asn Asp Phe Arg His Val Leu 225 230 235 240 cag cag gag ttg aag cgc cct gac ctg ctt cca ttg gac aat gaa tgc 768Gln Gln Glu Leu Lys Arg Pro Asp Leu Leu Pro Leu Asp Asn Glu Cys 245 250 255 ctt gag aag cga gtg ttc cat cgt ctc atc agc gga atg cat gcg tct 816Leu Glu Lys Arg Val Phe His Arg Leu Ile Ser Gly Met His Ala Ser 260 265 270 atc tcg acc cac ctt tgc tgg gac tac cta aac cag acg acg gga caa 864Ile Ser Thr His Leu Cys Trp Asp Tyr Leu Asn Gln Thr Thr Gly Gln 275 280 285 tgg cat cct aac ctt caa tgc ttc aaa gat cgt ctc cac gat cac ccc 912Trp His Pro Asn Leu Gln Cys Phe Lys Asp Arg Leu His Asp His Pro 290 295 300 gag cgc atc tcg aac ctg tac ttc aac tac gcg ctg gtc tcg cgc gcc 960Glu Arg Ile Ser Asn Leu Tyr Phe Asn Tyr Ala Leu Val Ser Arg Ala 305 310 315 320 gtg gcg aag ctg cag aaa cac cta cac aac tac aac tac tgc gtc ggt 1008Val Ala Lys Leu Gln Lys His Leu His Asn Tyr Asn Tyr Cys Val Gly 325 330 335 gat ccg gtc cag gat gcc atg act agg gag aag gtc tcc aag ttg acc 1056Asp Pro Val Gln Asp Ala Met Thr Arg Glu Lys Val Ser Lys Leu Thr 340 345 350 tcg acc ttg gct gac cgc cct caa att ttc gac gag aac gtc atg ttc 1104Ser Thr Leu Ala Asp Arg Pro Gln Ile Phe Asp Glu Asn Val Met Phe 355 360 365 cag gat ccc agc tcc gct ggc ctg aag gaa gac ttc cgc aac cga ttc 1152Gln Asp Pro Ser Ser Ala Gly Leu Lys Glu Asp Phe Arg Asn Arg Phe 370 375 380 cgc aac gtc agt cgc ctg atg gac tgc gtc ggg tgc gac aaa tgc cgc 1200Arg Asn Val Ser Arg Leu Met Asp Cys Val Gly Cys Asp Lys Cys Arg 385 390 395 400 ctc tgg ggc aag ctc cag gtc aac gga tat ggc acc gct ctg aaa gtg 1248Leu Trp Gly Lys Leu Gln Val Asn Gly Tyr Gly Thr Ala Leu Lys Val 405 410 415 ctg ttc gag tac gac gag act aag aac ggc gag aac ccg ttg ctg cgc 1296Leu Phe Glu Tyr Asp Glu Thr Lys Asn Gly Glu Asn Pro Leu Leu Arg 420 425 430 cgg act gag ctg gtg gca ctg atc aat acc ctt ggt cgc att tct cac 1344Arg Thr Glu Leu Val Ala Leu Ile Asn Thr Leu Gly Arg Ile Ser His 435 440 445 agc att gcc gcc gtc cgg agt ttc cac cgg gcc atg gat gtg ggc gat 1392Ser Ile Ala Ala Val Arg Ser Phe His Arg Ala Met Asp Val Gly Asp 450 455 460 ggg gag gtc ttc acc atc ccc gcg agc att gcg tcc aag gag cgc ggt 1440Gly Glu Val Phe Thr Ile Pro Ala Ser Ile Ala Ser Lys Glu Arg Gly 465 470 475 480 ggc aag aag aag acc cga cga ctt ctc aaa gac ggt ggc tca acc ttc 1488Gly Lys Lys Lys Thr Arg Arg Leu Leu Lys Asp Gly Gly Ser Thr Phe 485 490 495 tat tat gag gat ggc gat gat gac aac ttt gtc tac atc acc gag aaa 1536Tyr Tyr Glu Asp Gly Asp Asp Asp Asn Phe Val Tyr Ile Thr Glu Lys 500 505 510 ctt ccg tgg gag aag gtc cgg gta cgc cgc gac acg gat acg gtc tgg 1584Leu Pro Trp Glu Lys Val Arg Val Arg Arg Asp Thr Asp Thr Val Trp 515 520 525 gat gat att aag gcc gag ttt tct atg atc tgg gac att tac gtc tat 1632Asp Asp Ile Lys Ala Glu Phe Ser Met Ile Trp Asp Ile Tyr Val Tyr 530 535 540 gtg ctg aag agc tgg gtc aat gca cca aag aca ttc ttc gag atc gcc 1680Val Leu Lys Ser Trp Val Asn Ala Pro Lys Thr Phe Phe Glu Ile Ala 545 550 555 560 gtc ctg gag gtt gct cgg gta tgg aac tac tgg ctg ggt ctg cct gtg 1728Val Leu Glu Val Ala Arg Val Trp Asn Tyr Trp Leu Gly Leu Pro Val 565 570 575 ccg cca cgg tcc tgg agg atc cag ctt ccc aag cga ccc acc cct cca 1776Pro Pro Arg Ser Trp Arg Ile Gln Leu Pro Lys Arg Pro Thr Pro Pro 580 585 590 aca ccc ccg acc cat gag gag ctc tag 1803Thr Pro Pro Thr His Glu Glu Leu 595 600 36600PRTAspergillus niger 36Met Arg Ser Ala Ala Lys Phe Phe Tyr Leu Ala Val Phe Ala Leu Ser 1 5 10 15 Arg Leu Ser Asn Ala Glu Thr Gly Leu His His Asn Gln Asp Lys Cys 20 25 30 Ala Ile Asp Pro Thr Ala Met Val Ser Asp Ala Cys Val Ser Tyr Ala 35 40 45 Thr Ile Asp His Leu Asn Asp Gln Val Tyr Thr Leu Leu Gln Ser Ile 50 55 60 Thr Gln Asp Thr Asp Phe Phe Ser Tyr Tyr Arg Leu Asn Leu Phe Asn 65 70 75 80 Lys Val Cys Pro Phe Trp Ser Asp Ala Asn Ser Met Cys Gly Asn Ile 85 90 95 Ala Cys Ser Val Asn Thr Ile Glu Ser Glu Asp Asp Ile Pro Leu Thr 100 105 110 Trp Arg Ala Glu Glu Leu Ser Lys Leu Glu Gly Pro Lys Ala Gly His 115 120 125 Pro Gly Arg Asn Gln Arg Lys Glu Arg Pro Leu Asn Arg Pro Leu Gln 130 135 140 Gly Met Leu Gly Glu Asn Val Gly Glu Ser Cys Val Val Glu Tyr Asp 145 150 155 160 Asp Glu Cys Asp Glu Arg Asp Tyr Cys Val Pro Glu Asp Glu Gly Ala 165 170 175 Ser Gly Lys Gly Asp Tyr Val Ser Leu Val Asp Asn Pro Glu Arg Phe 180 185 190 Thr Gly Tyr Ala Gly Met Gly Ala His Gln Val Trp Asp Ala Ile Tyr 195 200 205 Arg Glu Asn Cys Phe Leu Lys Pro Val Pro Glu Leu Ser Pro Val Thr 210 215 220 Pro Gln Leu Gly Gly Leu Gln Ala Val Asn Asp Phe Arg His Val Leu 225 230 235 240 Gln Gln Glu Leu Lys Arg Pro Asp Leu Leu Pro Leu Asp Asn Glu Cys 245 250 255 Leu Glu Lys Arg Val Phe His Arg Leu Ile Ser Gly Met His Ala Ser 260 265 270 Ile Ser Thr His Leu Cys Trp Asp Tyr Leu Asn Gln Thr Thr Gly Gln 275 280 285 Trp His Pro Asn Leu Gln Cys Phe Lys Asp Arg Leu His Asp His Pro 290 295 300 Glu Arg Ile Ser Asn Leu Tyr Phe Asn Tyr Ala Leu Val Ser Arg Ala 305 310 315 320 Val Ala Lys Leu Gln Lys His Leu His Asn Tyr Asn Tyr Cys Val Gly 325 330 335 Asp Pro Val Gln Asp Ala Met Thr Arg Glu Lys Val Ser Lys Leu Thr 340 345 350 Ser Thr Leu Ala Asp Arg Pro Gln Ile Phe Asp Glu Asn Val Met Phe 355 360 365 Gln Asp Pro Ser Ser Ala Gly Leu Lys Glu Asp Phe Arg Asn Arg Phe 370 375 380 Arg Asn Val Ser Arg Leu Met Asp Cys Val Gly Cys Asp Lys Cys Arg 385 390 395 400 Leu Trp Gly Lys Leu Gln Val Asn Gly Tyr Gly Thr Ala Leu Lys Val 405 410 415 Leu Phe Glu Tyr Asp Glu Thr Lys Asn Gly Glu Asn Pro Leu Leu Arg 420 425 430 Arg Thr Glu Leu Val Ala Leu Ile Asn

Thr Leu Gly Arg Ile Ser His 435 440 445 Ser Ile Ala Ala Val Arg Ser Phe His Arg Ala Met Asp Val Gly Asp 450 455 460 Gly Glu Val Phe Thr Ile Pro Ala Ser Ile Ala Ser Lys Glu Arg Gly 465 470 475 480 Gly Lys Lys Lys Thr Arg Arg Leu Leu Lys Asp Gly Gly Ser Thr Phe 485 490 495 Tyr Tyr Glu Asp Gly Asp Asp Asp Asn Phe Val Tyr Ile Thr Glu Lys 500 505 510 Leu Pro Trp Glu Lys Val Arg Val Arg Arg Asp Thr Asp Thr Val Trp 515 520 525 Asp Asp Ile Lys Ala Glu Phe Ser Met Ile Trp Asp Ile Tyr Val Tyr 530 535 540 Val Leu Lys Ser Trp Val Asn Ala Pro Lys Thr Phe Phe Glu Ile Ala 545 550 555 560 Val Leu Glu Val Ala Arg Val Trp Asn Tyr Trp Leu Gly Leu Pro Val 565 570 575 Pro Pro Arg Ser Trp Arg Ile Gln Leu Pro Lys Arg Pro Thr Pro Pro 580 585 590 Thr Pro Pro Thr His Glu Glu Leu 595 600 372977DNAAspergillus niger 37gtggttgact tgggtgactt acgtacctgg cgccaagccc cagaagagac caccttgcga 60agctctcgac actcttcaat tcattccccc gttcacgaca cctcccaacc cttccctctc 120atcagctccc gggacaggtc gagggctgtt tgctcatctt ctttcccatt ggatcttgat 180tcttttctcc cctggccatt atggccgaat cacccctcga tgtcctcttg aagggtaact 240ccggcagaac aacacgcggc cttctgcgga ttatcattct agctaccatt gccgctgctg 300ctgtgtccag tcgtctgttc agtgtgatcc gtatgtctct taacttatat gatcgggtca 360attgaattat cgttactgac agttgccctc taggattcga gagtattatc cacgagtgta 420tgttttgata ccgatcctcc cgtctatacc ttcaagctac cgcggattcg gcattcgaca 480ccgtgtgcag ccaacttcac ttccaaaacc aatcaatgct aatgtagctc actcctacag 540tcgacccctg gttcaacttc cgcgcaacaa aatacctggt ctcccatggc tttgagagct 600tctgggactg gttcgacgac cgtacgttcc tcttgccaag cctgccttaa cctacatata 660ctgatctaag atgctgctca ggaacatggc accctctggg acgtgtcact ggtggcacgc 720tataccccgg tctcatggtg accagcggtg ttatttacca cgtcttgcgg ttcctcacta 780tccctgtcga catccgtaac atctgtgtct tgcttgcccc gggtttctcc ggtttgaccg 840cgctggcaat gtacttcctg actcgcgaga tggcgacatc cccctccgct ggtctcctcg 900cggctgcttt catgggtatc gtccctggat acatctctcg ttccgtcgca ggcagctacg 960ataacgaggc catcgccatt ttcctgctgg tattcacctt cttcctgtgg atcaaggctg 1020ttaagaatgg ctccatcatg tggggttctc tggcggcctt gttctacggc tacatggtgt 1080ctgcctgggg tggttatgtc ttcatcacta acttgatccc tctgcacgtt ttcgttcttc 1140tgtgcatggg cagatacagc tcgcgtatct atatcagtta taccacttgg tatgctctgg 1200gaactctggc cagtatgcag attcctttcg tcggattcct gccgattcgc aacagtgacc 1260acatgtccgc acttggtatg tactctcatt aaccgtagtg aagagcgttt gtactgacct 1320tgccaggtgt cttcggcctc attcagctcg tggcctttgc tgacttcgtc cggggtttca 1380ttccgggcag gcacttccag agacttctga ccaccatgat catcgtcgta tttggcatcg 1440ctttcgtcgg actcgtcgtc ctcaccgtgt ccggagtgat cgccccttgg agtggtcgtt 1500tctactctct gtgggatacc ggctatgcca agatccacat ccctatcatt gcgtccgtct 1560ccgagcacca gcccactgct tggcccgcct tcttctttga cctgaacttc ttgatctggc 1620tcttccctgc cggtgtctac atgtgcttcc gggatctcaa ggatgagcac gtttttgtca 1680tcatctactc ggtgcttgcc agttacttcg caggtgtcat ggttcgtctg atgttgactt 1740tgacccctat tgtttgtgtt gcggctgctc tggccctctc caccatcctc gacacgtatg 1800tgttcgcaaa gaatggcccc aacccccgcg ccaaggcgaa cgacgacacc tcggacggtc 1860ttcgttccac caggaagccc gatgttggtg tcacgtccta cctgtccaag gctgttatga 1920cttcctccgt tgtcatctat cttcttctct tcgttgcgca ctgcacctgg gtcacctcga 1980acgcatactc ctccccgtct gtggttctcg caagccgctt gcctgatgga agccagcaca 2040tcatcgacga ctaccgtgag gcgtactact ggcttcgtca gaacaccgag cacaacgcca 2100agatcatgtc ctggtgggat tacggctacc agattggtgg tatggcggac cgccctaccc 2160tggttgacaa caacacgtgg aacaacaccc acattgccac tgtcggtaag gcgatgagct 2220ctcgtgagga agtcagttac cccatcctcc gtcagcacga tgttgattac gtgctggtgg 2280tgtttggtgg attgctgggc tactccggtg acgacatcaa caagttcctg tggatggtcc 2340gtatcgctga aggtatctgg cccgacgagg tcaaggagcg tgacttcttc actgcccggg 2400gtgaataccg ggttgatgac ggagccaccc cgactatgcg taacagcttg atgtaagatt 2460tacccctgct tgccatggca gtgatatgat actaacagcg acccaggtac aaaatgtcct 2520actacaactt caactcgttg ttcggtcccg gccaggccgt tgaccgcgtg cgtggatcga 2580gactccccgc ggaaggccct cagctgaaca cgctcgagga ggcattcacc agtgagaact 2640ggatcatccg catctacaag gtcaaggatc tcgacaacct tggccgtgac cacaacaacg 2700cggtggcctt tgacaagggc cacaagcgca agcgcgctac caagcgcaag ggccctcgtg 2760tcctgcggac cgagtaaagg tccaggtgtc gttgatagat accaggttgg gtgtaaaatt 2820gatcccttct tcttcttcat ttcttagcat gtcattttca attccatgtt cttgtacgtg 2880tcatcaccat ataggcggaa tcatagaagt tgtccatctg gttagaggct gtagactgta 2940cattttaccc caagcaccaa atatgaacct tgagtga 2977382226DNAAspergillus nigerCDS(1)..(2226) 38atg gcc gaa tca ccc ctc gat gtc ctc ttg aag ggt aac tcc ggc aga 48Met Ala Glu Ser Pro Leu Asp Val Leu Leu Lys Gly Asn Ser Gly Arg 1 5 10 15 aca aca cgc ggc ctt ctg cgg att atc att cta gct acc att gcc gct 96Thr Thr Arg Gly Leu Leu Arg Ile Ile Ile Leu Ala Thr Ile Ala Ala 20 25 30 gct gct gtg tcc agt cgt ctg ttc agt gtg atc cga ttc gag agt att 144Ala Ala Val Ser Ser Arg Leu Phe Ser Val Ile Arg Phe Glu Ser Ile 35 40 45 atc cac gag ttc gac ccc tgg ttc aac ttc cgc gca aca aaa tac ctg 192Ile His Glu Phe Asp Pro Trp Phe Asn Phe Arg Ala Thr Lys Tyr Leu 50 55 60 gtc tcc cat ggc ttt gag agc ttc tgg gac tgg ttc gac gac cga aca 240Val Ser His Gly Phe Glu Ser Phe Trp Asp Trp Phe Asp Asp Arg Thr 65 70 75 80 tgg cac cct ctg gga cgt gtc act ggt ggc acg cta tac ccc ggt ctc 288Trp His Pro Leu Gly Arg Val Thr Gly Gly Thr Leu Tyr Pro Gly Leu 85 90 95 atg gtg acc agc ggt gtt att tac cac gtc ttg cgg ttc ctc act atc 336Met Val Thr Ser Gly Val Ile Tyr His Val Leu Arg Phe Leu Thr Ile 100 105 110 cct gtc gac atc cgt aac atc tgt gtc ttg ctt gcc ccg ggt ttc tcc 384Pro Val Asp Ile Arg Asn Ile Cys Val Leu Leu Ala Pro Gly Phe Ser 115 120 125 ggt ttg acc gcg ctg gca atg tac ttc ctg act cgc gag atg gcg aca 432Gly Leu Thr Ala Leu Ala Met Tyr Phe Leu Thr Arg Glu Met Ala Thr 130 135 140 tcc ccc tcc gct ggt ctc ctc gcg gct gct ttc atg ggt atc gtc cct 480Ser Pro Ser Ala Gly Leu Leu Ala Ala Ala Phe Met Gly Ile Val Pro 145 150 155 160 gga tac atc tct cgt tcc gtc gca ggc agc tac gat aac gag gcc atc 528Gly Tyr Ile Ser Arg Ser Val Ala Gly Ser Tyr Asp Asn Glu Ala Ile 165 170 175 gcc att ttc ctg ctg gta ttc acc ttc ttc ctg tgg atc aag gct gtt 576Ala Ile Phe Leu Leu Val Phe Thr Phe Phe Leu Trp Ile Lys Ala Val 180 185 190 aag aat ggc tcc atc atg tgg ggt tct ctg gcg gcc ttg ttc tac ggc 624Lys Asn Gly Ser Ile Met Trp Gly Ser Leu Ala Ala Leu Phe Tyr Gly 195 200 205 tac atg gtg tct gcc tgg ggt ggt tat gtc ttc atc act aac ttg atc 672Tyr Met Val Ser Ala Trp Gly Gly Tyr Val Phe Ile Thr Asn Leu Ile 210 215 220 cct ctg cac gtt ttc gtt ctt ctg tgc atg ggc aga tac agc tcg cgt 720Pro Leu His Val Phe Val Leu Leu Cys Met Gly Arg Tyr Ser Ser Arg 225 230 235 240 atc tat atc agt tat acc act tgg tat gct ctg gga act ctg gcc agt 768Ile Tyr Ile Ser Tyr Thr Thr Trp Tyr Ala Leu Gly Thr Leu Ala Ser 245 250 255 atg cag att cct ttc gtc gga ttc ctg ccg att cgc aac agt gac cac 816Met Gln Ile Pro Phe Val Gly Phe Leu Pro Ile Arg Asn Ser Asp His 260 265 270 atg tcc gca ctt ggt gtc ttc ggc ctc att cag ctc gtg gcc ttt gct 864Met Ser Ala Leu Gly Val Phe Gly Leu Ile Gln Leu Val Ala Phe Ala 275 280 285 gac ttc gtc cgg ggt ttc att ccg ggc agg cac ttc cag aga ctt ctg 912Asp Phe Val Arg Gly Phe Ile Pro Gly Arg His Phe Gln Arg Leu Leu 290 295 300 acc acc atg atc atc gtc gta ttt ggc atc gct ttc gtc gga ctc gtc 960Thr Thr Met Ile Ile Val Val Phe Gly Ile Ala Phe Val Gly Leu Val 305 310 315 320 gtc ctc acc gtg tcc gga gtg atc gcc cct tgg agt ggt cgt ttc tac 1008Val Leu Thr Val Ser Gly Val Ile Ala Pro Trp Ser Gly Arg Phe Tyr 325 330 335 tct ctg tgg gat acc ggc tat gcc aag atc cac atc cct atc att gcg 1056Ser Leu Trp Asp Thr Gly Tyr Ala Lys Ile His Ile Pro Ile Ile Ala 340 345 350 tcc gtc tcc gag cac cag ccc act gct tgg ccc gcc ttc ttc ttt gac 1104Ser Val Ser Glu His Gln Pro Thr Ala Trp Pro Ala Phe Phe Phe Asp 355 360 365 ctg aac ttc ttg atc tgg ctc ttc cct gcc ggt gtc tac atg tgc ttc 1152Leu Asn Phe Leu Ile Trp Leu Phe Pro Ala Gly Val Tyr Met Cys Phe 370 375 380 cgg gat ctc aag gat gag cac gtt ttt gtc atc atc tac tcg gtg ctt 1200Arg Asp Leu Lys Asp Glu His Val Phe Val Ile Ile Tyr Ser Val Leu 385 390 395 400 gcc agt tac ttc gca ggt gtc atg gtt cgt ctg atg ttg act ttg acc 1248Ala Ser Tyr Phe Ala Gly Val Met Val Arg Leu Met Leu Thr Leu Thr 405 410 415 cct att gtt tgt gtt gcg gct gct ctg gcc ctc tcc acc atc ctc gac 1296Pro Ile Val Cys Val Ala Ala Ala Leu Ala Leu Ser Thr Ile Leu Asp 420 425 430 acg tat gtg ttc gca aag aat ggc ccc aac ccc cgc gcc aag gcg aac 1344Thr Tyr Val Phe Ala Lys Asn Gly Pro Asn Pro Arg Ala Lys Ala Asn 435 440 445 gac gac acc tcg gac ggt ctt cgt tcc acc agg aag ccc gat gtt ggt 1392Asp Asp Thr Ser Asp Gly Leu Arg Ser Thr Arg Lys Pro Asp Val Gly 450 455 460 gtc acg tcc tac ctg tcc aag gct gtt atg act tcc tcc gtt gtc atc 1440Val Thr Ser Tyr Leu Ser Lys Ala Val Met Thr Ser Ser Val Val Ile 465 470 475 480 tat ctt ctt ctc ttc gtt gcg cac tgc acc tgg gtc acc tcg aac gca 1488Tyr Leu Leu Leu Phe Val Ala His Cys Thr Trp Val Thr Ser Asn Ala 485 490 495 tac tcc tcc ccg tct gtg gtt ctc gca agc cgc ttg cct gat gga agc 1536Tyr Ser Ser Pro Ser Val Val Leu Ala Ser Arg Leu Pro Asp Gly Ser 500 505 510 cag cac atc atc gac gac tac cgt gag gcg tac tac tgg ctt cgt cag 1584Gln His Ile Ile Asp Asp Tyr Arg Glu Ala Tyr Tyr Trp Leu Arg Gln 515 520 525 aac acc gag cac aac gcc aag atc atg tcc tgg tgg gat tac ggc tac 1632Asn Thr Glu His Asn Ala Lys Ile Met Ser Trp Trp Asp Tyr Gly Tyr 530 535 540 cag att ggt ggt atg gcg gac cgc cct acc ctg gtt gac aac aac acg 1680Gln Ile Gly Gly Met Ala Asp Arg Pro Thr Leu Val Asp Asn Asn Thr 545 550 555 560 tgg aac aac acc cac att gcc act gtc ggt aag gcg atg agc tct cgt 1728Trp Asn Asn Thr His Ile Ala Thr Val Gly Lys Ala Met Ser Ser Arg 565 570 575 gag gaa gtc agt tac ccc atc ctc cgt cag cac gat gtt gat tac gtg 1776Glu Glu Val Ser Tyr Pro Ile Leu Arg Gln His Asp Val Asp Tyr Val 580 585 590 ctg gtg gtg ttt ggt gga ttg ctg ggc tac tcc ggt gac gac atc aac 1824Leu Val Val Phe Gly Gly Leu Leu Gly Tyr Ser Gly Asp Asp Ile Asn 595 600 605 aag ttc ctg tgg atg gtc cgt atc gct gaa ggt atc tgg ccc gac gag 1872Lys Phe Leu Trp Met Val Arg Ile Ala Glu Gly Ile Trp Pro Asp Glu 610 615 620 gtc aag gag cgt gac ttc ttc act gcc cgg ggt gaa tac cgg gtt gat 1920Val Lys Glu Arg Asp Phe Phe Thr Ala Arg Gly Glu Tyr Arg Val Asp 625 630 635 640 gac gga gcc acc ccg act atg cgt aac agc ttg atg tac aaa atg tcc 1968Asp Gly Ala Thr Pro Thr Met Arg Asn Ser Leu Met Tyr Lys Met Ser 645 650 655 tac tac aac ttc aac tcg ttg ttc ggt ccc ggc cag gcc gtt gac cgc 2016Tyr Tyr Asn Phe Asn Ser Leu Phe Gly Pro Gly Gln Ala Val Asp Arg 660 665 670 gtg cgt gga tcg aga ctc ccc gcg gaa ggc cct cag ctg aac acg ctc 2064Val Arg Gly Ser Arg Leu Pro Ala Glu Gly Pro Gln Leu Asn Thr Leu 675 680 685 gag gag gca ttc acc agt gag aac tgg atc atc cgc atc tac aag gtc 2112Glu Glu Ala Phe Thr Ser Glu Asn Trp Ile Ile Arg Ile Tyr Lys Val 690 695 700 aag gat ctc gac aac ctt ggc cgt gac cac aac aac gcg gtg gcc ttt 2160Lys Asp Leu Asp Asn Leu Gly Arg Asp His Asn Asn Ala Val Ala Phe 705 710 715 720 gac aag ggc cac aag cgc aag cgc gct acc aag cgc aag ggc cct cgt 2208Asp Lys Gly His Lys Arg Lys Arg Ala Thr Lys Arg Lys Gly Pro Arg 725 730 735 gtc ctg cgg acc gag taa 2226Val Leu Arg Thr Glu 740 39741PRTAspergillus niger 39Met Ala Glu Ser Pro Leu Asp Val Leu Leu Lys Gly Asn Ser Gly Arg 1 5 10 15 Thr Thr Arg Gly Leu Leu Arg Ile Ile Ile Leu Ala Thr Ile Ala Ala 20 25 30 Ala Ala Val Ser Ser Arg Leu Phe Ser Val Ile Arg Phe Glu Ser Ile 35 40 45 Ile His Glu Phe Asp Pro Trp Phe Asn Phe Arg Ala Thr Lys Tyr Leu 50 55 60 Val Ser His Gly Phe Glu Ser Phe Trp Asp Trp Phe Asp Asp Arg Thr 65 70 75 80 Trp His Pro Leu Gly Arg Val Thr Gly Gly Thr Leu Tyr Pro Gly Leu 85 90 95 Met Val Thr Ser Gly Val Ile Tyr His Val Leu Arg Phe Leu Thr Ile 100 105 110 Pro Val Asp Ile Arg Asn Ile Cys Val Leu Leu Ala Pro Gly Phe Ser 115 120 125 Gly Leu Thr Ala Leu Ala Met Tyr Phe Leu Thr Arg Glu Met Ala Thr 130 135 140 Ser Pro Ser Ala Gly Leu Leu Ala Ala Ala Phe Met Gly Ile Val Pro 145 150 155 160 Gly Tyr Ile Ser Arg Ser Val Ala Gly Ser Tyr Asp Asn Glu Ala Ile 165 170 175 Ala Ile Phe Leu Leu Val Phe Thr Phe Phe Leu Trp Ile Lys Ala Val 180 185 190 Lys Asn Gly Ser Ile Met Trp Gly Ser Leu Ala Ala Leu Phe Tyr Gly 195 200 205 Tyr Met Val Ser Ala Trp Gly Gly Tyr Val Phe Ile Thr Asn Leu Ile 210 215 220 Pro Leu His Val Phe Val Leu Leu Cys Met Gly Arg Tyr Ser Ser Arg 225 230 235 240 Ile Tyr Ile Ser Tyr Thr Thr Trp Tyr Ala Leu Gly Thr Leu Ala Ser 245 250 255 Met Gln Ile Pro Phe Val Gly Phe Leu Pro Ile Arg Asn Ser Asp His 260 265 270 Met Ser Ala Leu Gly Val Phe Gly Leu Ile Gln Leu Val Ala Phe Ala 275 280 285 Asp Phe Val Arg Gly Phe Ile Pro Gly Arg His Phe Gln Arg Leu Leu 290 295 300 Thr Thr Met Ile Ile Val Val Phe Gly Ile Ala Phe Val Gly Leu Val 305 310 315 320 Val Leu Thr Val Ser Gly Val Ile Ala Pro Trp Ser Gly Arg Phe Tyr 325 330 335 Ser Leu Trp Asp Thr Gly Tyr Ala Lys Ile His Ile Pro Ile Ile Ala 340 345 350 Ser Val Ser Glu His Gln Pro Thr Ala Trp Pro Ala Phe Phe Phe Asp 355 360 365 Leu Asn Phe Leu Ile Trp Leu Phe Pro Ala Gly Val Tyr Met Cys Phe 370 375 380 Arg Asp Leu Lys Asp Glu His Val Phe Val Ile Ile Tyr Ser Val Leu 385 390 395 400 Ala Ser Tyr Phe Ala Gly Val Met Val Arg Leu Met Leu Thr Leu Thr 405 410 415 Pro Ile Val Cys Val Ala Ala Ala Leu Ala Leu Ser Thr Ile Leu Asp 420 425 430 Thr Tyr Val Phe Ala Lys Asn Gly Pro Asn Pro Arg Ala Lys Ala Asn 435 440 445 Asp Asp Thr Ser Asp Gly Leu Arg Ser Thr Arg Lys Pro Asp Val Gly 450 455 460 Val Thr Ser Tyr Leu Ser Lys Ala Val Met Thr Ser Ser Val Val Ile 465 470 475 480 Tyr Leu Leu Leu Phe Val Ala His Cys Thr Trp Val Thr Ser Asn Ala 485 490

495 Tyr Ser Ser Pro Ser Val Val Leu Ala Ser Arg Leu Pro Asp Gly Ser 500 505 510 Gln His Ile Ile Asp Asp Tyr Arg Glu Ala Tyr Tyr Trp Leu Arg Gln 515 520 525 Asn Thr Glu His Asn Ala Lys Ile Met Ser Trp Trp Asp Tyr Gly Tyr 530 535 540 Gln Ile Gly Gly Met Ala Asp Arg Pro Thr Leu Val Asp Asn Asn Thr 545 550 555 560 Trp Asn Asn Thr His Ile Ala Thr Val Gly Lys Ala Met Ser Ser Arg 565 570 575 Glu Glu Val Ser Tyr Pro Ile Leu Arg Gln His Asp Val Asp Tyr Val 580 585 590 Leu Val Val Phe Gly Gly Leu Leu Gly Tyr Ser Gly Asp Asp Ile Asn 595 600 605 Lys Phe Leu Trp Met Val Arg Ile Ala Glu Gly Ile Trp Pro Asp Glu 610 615 620 Val Lys Glu Arg Asp Phe Phe Thr Ala Arg Gly Glu Tyr Arg Val Asp 625 630 635 640 Asp Gly Ala Thr Pro Thr Met Arg Asn Ser Leu Met Tyr Lys Met Ser 645 650 655 Tyr Tyr Asn Phe Asn Ser Leu Phe Gly Pro Gly Gln Ala Val Asp Arg 660 665 670 Val Arg Gly Ser Arg Leu Pro Ala Glu Gly Pro Gln Leu Asn Thr Leu 675 680 685 Glu Glu Ala Phe Thr Ser Glu Asn Trp Ile Ile Arg Ile Tyr Lys Val 690 695 700 Lys Asp Leu Asp Asn Leu Gly Arg Asp His Asn Asn Ala Val Ala Phe 705 710 715 720 Asp Lys Gly His Lys Arg Lys Arg Ala Thr Lys Arg Lys Gly Pro Arg 725 730 735 Val Leu Arg Thr Glu 740 401546DNAAspergillus niger 40ttaccaagtg tcctctttga cccgctcggt ctccgcagcg cttccgcctt ccacgccggg 60agctcatacg actatatcta cactccgccc agcaacaacc ccttccctcg ctgaacggac 120aggaccattt gctattgaca agtaacattg tcaatcgccc tgcttctact tctccctaaa 180ccaaaaccac accatccatc atggtccgtc tcagcaatct cgtgagctgc ctcggcctgg 240cctccgcggt caccgcagca gtggtcgatc tcgtccccaa gaacttcgac gacgtcgtcc 300tcaagtccgg caagcccgct ctggttgaat tcttcgctcc ctggtgcggc cactgcaaga 360acctcgcgcc cgtgtatgaa gagctgggcc aggcattcgc ccatgcctcc gacaaggtca 420ccgtcggcaa ggttgatgcg gacgagcacc gcgacttggg ccgcaagttc ggtgtccagg 480gattccccac gctaaagtgg ttcgacggaa agagtgacga gccggaggat tacaagggtg 540gtcgtgattt ggagagtctg tcttcgttca tctctgagaa gacgggcgtc aagccccgtg 600gtcctaagaa ggagcccagc aaggtggaga tgctgaacga cgcgactttc aagggcgctg 660ttggtggcga taatgatgtt ctggttgcgt tcaccgcgcc gtggtgtgga cgtgagtatc 720ctcgtttcat cgttcccgct ctagaagcaa atcactaact acggcctttt aaaacagact 780gcaagaacct cgctcctacc tgggaagccc tggccaacga cttcgtcctc gagcccaacg 840ttgtgatcgc caaggtcgac gccgacgctg agaacggcaa ggccaccgcc agagagcagg 900gcgtgtccgg ataccccacc atcaagttct tccccaaggg ctctacggaa tctgttccct 960atgagggtgc ccgctctgag caggccttca ttgacttcct caacgagaag accggcaccc 1020accgtaccgt tggcggcgga ctcgacacca aggccggcac cattgctagc ctggacgagc 1080tgattgccag cacttctgct gctgacctgg ccgccgcagt caagaaggct gctacggagc 1140ttaaggacaa gtacgctcag tactacgtca aggttgcgga caagctgagc cagaacgccg 1200agtatgccgc taaggagctt gctcgtctgg agaagatcct ggccaagggt ggatcggccc 1260ctgagaaggt ggatgacctt atctcccgca gcaacatcct tcgcaagttt gttggtgagg 1320agaaggaggc caaggatgag ctgtagatat tgtatggatt atgacttgtt tagctagggt 1380ataggcacct agtttctgtt actctgtatg atatcaagag gcagttatga gaatgctatc 1440gatgcgaaca gtaaaccatc catttcccat tcccatgtat gtatacacaa gaacataaga 1500gtatatagta gatgtaaatt gacagtaaaa agcgtatctt cacatt 1546411080DNAAspergillus nigerCDS(1)..(1080) 41atg gtc cgt ctc agc aat ctc gtg agc tgc ctc ggc ctg gcc tcc gcg 48Met Val Arg Leu Ser Asn Leu Val Ser Cys Leu Gly Leu Ala Ser Ala 1 5 10 15 gtc acc gca gca gtg gtc gat ctc gtc ccc aag aac ttc gac gac gtc 96Val Thr Ala Ala Val Val Asp Leu Val Pro Lys Asn Phe Asp Asp Val 20 25 30 gtc ctc aag tcc ggc aag ccc gct ctg gtt gaa ttc ttc gct ccc tgg 144Val Leu Lys Ser Gly Lys Pro Ala Leu Val Glu Phe Phe Ala Pro Trp 35 40 45 tgc ggc cac tgc aag aac ctc gcg ccc gtg tat gaa gag ctg ggc cag 192Cys Gly His Cys Lys Asn Leu Ala Pro Val Tyr Glu Glu Leu Gly Gln 50 55 60 gca ttc gcc cat gcc tcc gac aag gtc acc gtc ggc aag gtt gat gcg 240Ala Phe Ala His Ala Ser Asp Lys Val Thr Val Gly Lys Val Asp Ala 65 70 75 80 gac gag cac cgc gac ttg ggc cgc aag ttc ggt gtc cag gga ttc ccc 288Asp Glu His Arg Asp Leu Gly Arg Lys Phe Gly Val Gln Gly Phe Pro 85 90 95 acg cta aag tgg ttc gac gga aag agt gac gag ccg gag gat tac aag 336Thr Leu Lys Trp Phe Asp Gly Lys Ser Asp Glu Pro Glu Asp Tyr Lys 100 105 110 ggt ggt cgt gat ttg gag agt ctg tct tcg ttc atc tct gag aag acg 384Gly Gly Arg Asp Leu Glu Ser Leu Ser Ser Phe Ile Ser Glu Lys Thr 115 120 125 ggc gtc aag ccc cgt ggt cct aag aag gag ccc agc aag gtg gag atg 432Gly Val Lys Pro Arg Gly Pro Lys Lys Glu Pro Ser Lys Val Glu Met 130 135 140 ctg aac gac gcg act ttc aag ggc gct gtt ggt ggc gat aat gat gtt 480Leu Asn Asp Ala Thr Phe Lys Gly Ala Val Gly Gly Asp Asn Asp Val 145 150 155 160 ctg gtt gcg ttc acc gcg ccg tgg tgt gga cac tgc aag aac ctc gct 528Leu Val Ala Phe Thr Ala Pro Trp Cys Gly His Cys Lys Asn Leu Ala 165 170 175 cct acc tgg gaa gcc ctg gcc aac gac ttc gtc ctc gag ccc aac gtt 576Pro Thr Trp Glu Ala Leu Ala Asn Asp Phe Val Leu Glu Pro Asn Val 180 185 190 gtg atc gcc aag gtc gac gcc gac gct gag aac ggc aag gcc acc gcc 624Val Ile Ala Lys Val Asp Ala Asp Ala Glu Asn Gly Lys Ala Thr Ala 195 200 205 aga gag cag ggc gtg tcc gga tac ccc acc atc aag ttc ttc ccc aag 672Arg Glu Gln Gly Val Ser Gly Tyr Pro Thr Ile Lys Phe Phe Pro Lys 210 215 220 ggc tct acg gaa tct gtt ccc tat gag ggt gcc cgc tct gag cag gcc 720Gly Ser Thr Glu Ser Val Pro Tyr Glu Gly Ala Arg Ser Glu Gln Ala 225 230 235 240 ttc att gac ttc ctc aac gag aag acc ggc acc cac cgt acc gtt ggc 768Phe Ile Asp Phe Leu Asn Glu Lys Thr Gly Thr His Arg Thr Val Gly 245 250 255 ggc gga ctc gac acc aag gcc ggc acc att gct agc ctg gac gag ctg 816Gly Gly Leu Asp Thr Lys Ala Gly Thr Ile Ala Ser Leu Asp Glu Leu 260 265 270 att gcc agc act tct gct gct gac ctg gcc gcc gca gtc aag aag gct 864Ile Ala Ser Thr Ser Ala Ala Asp Leu Ala Ala Ala Val Lys Lys Ala 275 280 285 gct acg gag ctt aag gac aag tac gct cag tac tac gtc aag gtt gcg 912Ala Thr Glu Leu Lys Asp Lys Tyr Ala Gln Tyr Tyr Val Lys Val Ala 290 295 300 gac aag ctg agc cag aac gcc gag tat gcc gct aag gag ctt gct cgt 960Asp Lys Leu Ser Gln Asn Ala Glu Tyr Ala Ala Lys Glu Leu Ala Arg 305 310 315 320 ctg gag aag atc ctg gcc aag ggt gga tcg gcc cct gag aag gtg gat 1008Leu Glu Lys Ile Leu Ala Lys Gly Gly Ser Ala Pro Glu Lys Val Asp 325 330 335 gac ctt atc tcc cgc agc aac atc ctt cgc aag ttt gtt ggt gag gag 1056Asp Leu Ile Ser Arg Ser Asn Ile Leu Arg Lys Phe Val Gly Glu Glu 340 345 350 aag gag gcc aag gat gag ctg tag 1080Lys Glu Ala Lys Asp Glu Leu 355 42359PRTAspergillus niger 42Met Val Arg Leu Ser Asn Leu Val Ser Cys Leu Gly Leu Ala Ser Ala 1 5 10 15 Val Thr Ala Ala Val Val Asp Leu Val Pro Lys Asn Phe Asp Asp Val 20 25 30 Val Leu Lys Ser Gly Lys Pro Ala Leu Val Glu Phe Phe Ala Pro Trp 35 40 45 Cys Gly His Cys Lys Asn Leu Ala Pro Val Tyr Glu Glu Leu Gly Gln 50 55 60 Ala Phe Ala His Ala Ser Asp Lys Val Thr Val Gly Lys Val Asp Ala 65 70 75 80 Asp Glu His Arg Asp Leu Gly Arg Lys Phe Gly Val Gln Gly Phe Pro 85 90 95 Thr Leu Lys Trp Phe Asp Gly Lys Ser Asp Glu Pro Glu Asp Tyr Lys 100 105 110 Gly Gly Arg Asp Leu Glu Ser Leu Ser Ser Phe Ile Ser Glu Lys Thr 115 120 125 Gly Val Lys Pro Arg Gly Pro Lys Lys Glu Pro Ser Lys Val Glu Met 130 135 140 Leu Asn Asp Ala Thr Phe Lys Gly Ala Val Gly Gly Asp Asn Asp Val 145 150 155 160 Leu Val Ala Phe Thr Ala Pro Trp Cys Gly His Cys Lys Asn Leu Ala 165 170 175 Pro Thr Trp Glu Ala Leu Ala Asn Asp Phe Val Leu Glu Pro Asn Val 180 185 190 Val Ile Ala Lys Val Asp Ala Asp Ala Glu Asn Gly Lys Ala Thr Ala 195 200 205 Arg Glu Gln Gly Val Ser Gly Tyr Pro Thr Ile Lys Phe Phe Pro Lys 210 215 220 Gly Ser Thr Glu Ser Val Pro Tyr Glu Gly Ala Arg Ser Glu Gln Ala 225 230 235 240 Phe Ile Asp Phe Leu Asn Glu Lys Thr Gly Thr His Arg Thr Val Gly 245 250 255 Gly Gly Leu Asp Thr Lys Ala Gly Thr Ile Ala Ser Leu Asp Glu Leu 260 265 270 Ile Ala Ser Thr Ser Ala Ala Asp Leu Ala Ala Ala Val Lys Lys Ala 275 280 285 Ala Thr Glu Leu Lys Asp Lys Tyr Ala Gln Tyr Tyr Val Lys Val Ala 290 295 300 Asp Lys Leu Ser Gln Asn Ala Glu Tyr Ala Ala Lys Glu Leu Ala Arg 305 310 315 320 Leu Glu Lys Ile Leu Ala Lys Gly Gly Ser Ala Pro Glu Lys Val Asp 325 330 335 Asp Leu Ile Ser Arg Ser Asn Ile Leu Arg Lys Phe Val Gly Glu Glu 340 345 350 Lys Glu Ala Lys Asp Glu Leu 355 432673DNAAspergillus niger 43gccaagaacg tcggcgaaat gaagctccgc aacgccaggt ggcccgttcc gcatcggggc 60cgagcaagca actccccccg agtctcaagt gctcatcttc actatcagct cctctcccct 120ctcttacttt atcggaagct cgagcacaga gtcgccatgc gcctctcact gttcacttct 180attcaacttt ctcacacaac atgaagaact ggtggctgtg gcgtttcctg ccgttggcgc 240ttcgtgagtc ctcttactgt ggagcttagc cgcatcacgc gatagcaact gacctccaag 300catcactttc cagtgcttct gcaggcgctt gcggatgaat acaatggaca gcacgacaca 360caaaaaccct taacagatgt tgttcctgaa tcatatgcac atgctgagtc ctcgagcggg 420cccgagggct ctgacgttct accgggacac ggtaagcggc cctgtgcatg acgccagcgc 480acccatcctt gctaacaact agtgatttga tgtgccaata gtacacgtcg aaaatgccct 540tcaaatcctc cgagagagca agatccccat tgtcgctcac gagaaaccgt ccggccttct 600ggggtacacc tggcattacg cccaagaagc cttccgactc ctatttatga atggaccaca 660gccggatgga acacacaagc aaaagctcga tccaaatgtt gcaaaggctg cgaatgaact 720taaggttgcg gcgcaagaac accaaaaccc cgatgcaatg ttcctcttag cggaaatgaa 780cttctacggc aacttcaccc acccgagaga tttcaagcag gcgtttcatt ggtaccaaac 840tctggcgtca tcgactggaa acagtacggc gcaatatatg cttgggttta tgtatgcaac 900gggtgtcggg ggtgcggtgg agcgcgacca ggctaaggcc ctgttatacc acacctttgc 960ggctgaagcg ggcaatacga agtcggaaat gaccctcgcg tatcgctacc acgctggaat 1020tggggctcct agagattgcg atcaagcgac ttactactat aagaaggtgg ctgataaggc 1080tattgaatac ttccgatcgg gaccgcccgg tggccataac atgatccgcg agtcctaccg 1140ttgggcggac gaagagggtg gtgtttatgg tgaaggcgct agtgtatcga ctgccgtacg 1200cgatggaacg cattcgagca cggaagccag cttggaagac gtcttggagt acctggattt 1260gatgtcgaga aagggcgaac tgaaggctac tttcagcttg ggcaagatgc attacgaagg 1320gggccgcggc ttgcctcgga atttccgaaa gtcgatgaat tacttccgac aggtcgccaa 1380gcggtattgg aataaagatg gatcggtgaa ccccaaccat cctgttggtg ttgaaaagct 1440cgcttcgaaa gcagcaggcc atattggcat gatgtacctg cgtggcgagg gggtggaaca 1500gaactttgca accgctcaga cttggtttag gcgtggactc gcgaatggtg atgctctctg 1560tcagcatgag ctaggactga tgtacctgca tggctatggt gtgacaccag atgcgttcag 1620agctgcatca caatttaagg ctgcggctga gcaggacttc ccggcggctg aaacgagact 1680gggtgccctg tttctagacc agggtgatgt ccagaccgcc acccgttatt tcgaactggc 1740tgcgcgctgg ggatggatgg aggccttcta ctacctggca gaattgtcca acaatggggt 1800tggtcggaaa cgacactgcg ggatggccgc gtcttactac aagatggtcg cagagcgggc 1860ggaagtcatc cattcatctt ttgaggaagc aaatacggcg tatgagaacg gagacaagga 1920acgggctctc attccggcgc tgatggctgc ggagcagggt tacgagcatg cacagtccaa 1980tgttgcgttc ctgctggacg agcagcggtc cttattcgcc attgacacta tcctcccagg 2040agctaagaag agcagaccgg ctttgctgcg gaatgcagcg ctggctctta tctattggac 2100acgttccgcc aaacaggcga acatcgactc cttgctcaag atgggcgatt actacctggc 2160gggcatggga attgctgcgg atgcggagaa ggcctcgacc tgctaccaca cagcagccga 2220agtgcactat agcgcacagg cgtactggaa tctgggatgg atgcatgaga atggcgttgc 2280ggtggaccaa gacttccaca tggccaagcg atactacgat ctagcgctgg agactagctc 2340cgaggcatat ctgcccgtga agctcagtct gcttaaactg cggatgcggg gatactggaa 2400ctggctcacg aacggagaca tcaaccctat ccgagaggaa gaaggtaagg aaccccaaca 2460tccttttcgc tgagaaaccc gaacaatcac acttacacac taatgcagaa gtgaggtcgc 2520atcgcacctt gaaggaattc atcgccactt ttatccagaa caacgaggaa gaagaggccg 2580ccttccgcgc ccagatgtac aaacaggacg aggaggacga actcatgtcg aataatcgcc 2640ttgacgacca ccgcgaagac ggctactatg atg 2673442070DNAAspergillus nigerCDS(1)..(2070) 44atg aag aac tgg tgg ctg tgg cgt ttc ctg ccg ttg gcg ctt cat gtt 48Met Lys Asn Trp Trp Leu Trp Arg Phe Leu Pro Leu Ala Leu His Val 1 5 10 15 gtt cct gaa tca tat gca cat gct gag tcc tcg agc ggg ccc gag ggc 96Val Pro Glu Ser Tyr Ala His Ala Glu Ser Ser Ser Gly Pro Glu Gly 20 25 30 tct gac gtt cta ccg gga cac gta cac gtc gaa aat gcc ctt caa atc 144Ser Asp Val Leu Pro Gly His Val His Val Glu Asn Ala Leu Gln Ile 35 40 45 ctc cga gag agc aag atc ccc att gtc gct cac gag aaa ccg tcc ggc 192Leu Arg Glu Ser Lys Ile Pro Ile Val Ala His Glu Lys Pro Ser Gly 50 55 60 ctt ctg ggg tac acc tgg cat tac gcc caa gaa gcc ttc cga ctc cta 240Leu Leu Gly Tyr Thr Trp His Tyr Ala Gln Glu Ala Phe Arg Leu Leu 65 70 75 80 ttt atg aat gga cca cag ccg gat gga aca cac aag caa aag ctc gat 288Phe Met Asn Gly Pro Gln Pro Asp Gly Thr His Lys Gln Lys Leu Asp 85 90 95 cca aat gtt gca aag gct gcg aat gaa ctt aag gtt gcg gcg caa gaa 336Pro Asn Val Ala Lys Ala Ala Asn Glu Leu Lys Val Ala Ala Gln Glu 100 105 110 cac caa aac ccc gat gca atg ttc ctc tta gcg gaa atg aac ttc tac 384His Gln Asn Pro Asp Ala Met Phe Leu Leu Ala Glu Met Asn Phe Tyr 115 120 125 ggc aac ttc acc cac ccg aga gat ttc aag cag gcg ttt cat tgg tac 432Gly Asn Phe Thr His Pro Arg Asp Phe Lys Gln Ala Phe His Trp Tyr 130 135 140 caa act ctg gcg tca tcg act gga aac agt acg gcg caa tat atg ctt 480Gln Thr Leu Ala Ser Ser Thr Gly Asn Ser Thr Ala Gln Tyr Met Leu 145 150 155 160 ggg ttt atg tat gca acg ggt gtc ggg ggt gcg gtg gag cgc gac cag 528Gly Phe Met Tyr Ala Thr Gly Val Gly Gly Ala Val Glu Arg Asp Gln 165 170 175 gct aag gcc ctg tta tac cac acc ttt gcg gct gaa gcg ggc aat acg 576Ala Lys Ala Leu Leu Tyr His Thr Phe Ala Ala Glu Ala Gly Asn Thr 180 185 190 aag tcg gaa atg acc ctc gcg tat cgc tac cac gct gga att ggg gct 624Lys Ser Glu Met Thr Leu Ala Tyr Arg Tyr His Ala Gly Ile Gly Ala 195 200 205 cct aga gat tgc gat caa gcg act tac tac tat aag aag gtg gct gat 672Pro Arg Asp Cys Asp Gln Ala Thr Tyr Tyr Tyr Lys Lys Val Ala Asp 210 215 220 aag gct att gaa tac ttc cga tcg gga ccg ccc ggt ggc cat aac atg 720Lys Ala Ile Glu Tyr Phe Arg Ser Gly Pro Pro Gly Gly His Asn Met 225 230 235 240 atc cgc gag tcc tac cgt tgg gcg gac gaa gag ggt ggt gtt tat ggt 768Ile Arg Glu Ser Tyr Arg Trp Ala Asp Glu Glu Gly Gly Val Tyr Gly 245 250 255 gaa ggc gct agt gta tcg act gcc gta cgc gat gga acg cat tcg agc 816Glu Gly Ala Ser Val Ser Thr Ala Val Arg Asp Gly Thr His Ser Ser 260 265 270 acg gaa gcc agc ttg gaa gac gtc ttg gag tac ctg gat ttg atg tcg 864Thr Glu Ala Ser Leu Glu Asp Val Leu Glu Tyr Leu Asp Leu Met Ser 275 280 285

aga aag ggc gaa ctg aag gct act ttc agc ttg ggc aag atg cat tac 912Arg Lys Gly Glu Leu Lys Ala Thr Phe Ser Leu Gly Lys Met His Tyr 290 295 300 gaa ggg ggc cgc ggc ttg cct cgg aat ttc cga aag tcg atg aat tac 960Glu Gly Gly Arg Gly Leu Pro Arg Asn Phe Arg Lys Ser Met Asn Tyr 305 310 315 320 ttc cga cag gtc gcc aag cgg tat tgg aat aaa gat gga tcg gtg aac 1008Phe Arg Gln Val Ala Lys Arg Tyr Trp Asn Lys Asp Gly Ser Val Asn 325 330 335 ccc aac cat cct gtt ggt gtt gaa aag ctc gct tcg aaa gca gca ggc 1056Pro Asn His Pro Val Gly Val Glu Lys Leu Ala Ser Lys Ala Ala Gly 340 345 350 cat att ggc atg atg tac ctg cgt ggc gag ggg gtg gaa cag aac ttt 1104His Ile Gly Met Met Tyr Leu Arg Gly Glu Gly Val Glu Gln Asn Phe 355 360 365 gca acc gct cag act tgg ttt agg cgt gga ctc gcg aat ggt gat gct 1152Ala Thr Ala Gln Thr Trp Phe Arg Arg Gly Leu Ala Asn Gly Asp Ala 370 375 380 ctc tgt cag cat gag cta gga ctg atg tac ctg cat ggc tat ggt gtg 1200Leu Cys Gln His Glu Leu Gly Leu Met Tyr Leu His Gly Tyr Gly Val 385 390 395 400 aca cca gat gcg ttc aga gct gca tca caa ttt aag gct gcg gct gag 1248Thr Pro Asp Ala Phe Arg Ala Ala Ser Gln Phe Lys Ala Ala Ala Glu 405 410 415 cag gac ttc ccg gcg gct gaa acg aga ctg ggt gcc ctg ttt cta gac 1296Gln Asp Phe Pro Ala Ala Glu Thr Arg Leu Gly Ala Leu Phe Leu Asp 420 425 430 cag ggt gat gtc cag acc gcc acc cgt tat ttc gaa ctg gct gcg cgc 1344Gln Gly Asp Val Gln Thr Ala Thr Arg Tyr Phe Glu Leu Ala Ala Arg 435 440 445 tgg gga tgg atg gag gcc ttc tac tac ctg gca gaa ttg tcc aac aat 1392Trp Gly Trp Met Glu Ala Phe Tyr Tyr Leu Ala Glu Leu Ser Asn Asn 450 455 460 ggg gtt ggt cgg aaa cga cac tgc ggg atg gcc gcg tct tac tac aag 1440Gly Val Gly Arg Lys Arg His Cys Gly Met Ala Ala Ser Tyr Tyr Lys 465 470 475 480 atg gtc gca gag cgg gcg gaa gtc atc cat tca tct ttt gag gaa gca 1488Met Val Ala Glu Arg Ala Glu Val Ile His Ser Ser Phe Glu Glu Ala 485 490 495 aat acg gcg tat gag aac gga gac aag gaa cgg gct ctc att ccg gcg 1536Asn Thr Ala Tyr Glu Asn Gly Asp Lys Glu Arg Ala Leu Ile Pro Ala 500 505 510 ctg atg gct gcg gag cag ggt tac gag cat gca cag tcc aat gtt gcg 1584Leu Met Ala Ala Glu Gln Gly Tyr Glu His Ala Gln Ser Asn Val Ala 515 520 525 ttc ctg ctg gac gag cag cgg tcc tta ttc gcc att gac act atc ctc 1632Phe Leu Leu Asp Glu Gln Arg Ser Leu Phe Ala Ile Asp Thr Ile Leu 530 535 540 cca gga gct aag aag agc aga ccg gct ttg ctg cgg aat gca gcg ctg 1680Pro Gly Ala Lys Lys Ser Arg Pro Ala Leu Leu Arg Asn Ala Ala Leu 545 550 555 560 gct ctt atc tat tgg aca cgt tcc gcc aaa cag gcg aac atc gac tcc 1728Ala Leu Ile Tyr Trp Thr Arg Ser Ala Lys Gln Ala Asn Ile Asp Ser 565 570 575 ttg ctc aag atg ggc gat tac tac ctg gcg ggc atg gga att gct gcg 1776Leu Leu Lys Met Gly Asp Tyr Tyr Leu Ala Gly Met Gly Ile Ala Ala 580 585 590 gat gcg gag aag gcc tcg acc tgc tac cac aca gca gcc gaa gtg cac 1824Asp Ala Glu Lys Ala Ser Thr Cys Tyr His Thr Ala Ala Glu Val His 595 600 605 tat agc gca cag gcg tac tgg aat ctg gga tgg atg cat gag aat ggc 1872Tyr Ser Ala Gln Ala Tyr Trp Asn Leu Gly Trp Met His Glu Asn Gly 610 615 620 gtt gcg gtg gac caa gac ttc cac atg gcc aag cga tac tac gat cta 1920Val Ala Val Asp Gln Asp Phe His Met Ala Lys Arg Tyr Tyr Asp Leu 625 630 635 640 gcg ctg gag act agc tcc gag gca tat ctg ccc gtg aag ctc agt ctg 1968Ala Leu Glu Thr Ser Ser Glu Ala Tyr Leu Pro Val Lys Leu Ser Leu 645 650 655 ctt aaa ctg cgg atg cgg gga tac tgg aac tgg ctc acg aac gga gac 2016Leu Lys Leu Arg Met Arg Gly Tyr Trp Asn Trp Leu Thr Asn Gly Asp 660 665 670 atc aac cct atc cga gag gaa gaa ggt aag gaa ccc caa cat cct ttt 2064Ile Asn Pro Ile Arg Glu Glu Glu Gly Lys Glu Pro Gln His Pro Phe 675 680 685 cgc tga 2070Arg 45689PRTAspergillus niger 45Met Lys Asn Trp Trp Leu Trp Arg Phe Leu Pro Leu Ala Leu His Val 1 5 10 15 Val Pro Glu Ser Tyr Ala His Ala Glu Ser Ser Ser Gly Pro Glu Gly 20 25 30 Ser Asp Val Leu Pro Gly His Val His Val Glu Asn Ala Leu Gln Ile 35 40 45 Leu Arg Glu Ser Lys Ile Pro Ile Val Ala His Glu Lys Pro Ser Gly 50 55 60 Leu Leu Gly Tyr Thr Trp His Tyr Ala Gln Glu Ala Phe Arg Leu Leu 65 70 75 80 Phe Met Asn Gly Pro Gln Pro Asp Gly Thr His Lys Gln Lys Leu Asp 85 90 95 Pro Asn Val Ala Lys Ala Ala Asn Glu Leu Lys Val Ala Ala Gln Glu 100 105 110 His Gln Asn Pro Asp Ala Met Phe Leu Leu Ala Glu Met Asn Phe Tyr 115 120 125 Gly Asn Phe Thr His Pro Arg Asp Phe Lys Gln Ala Phe His Trp Tyr 130 135 140 Gln Thr Leu Ala Ser Ser Thr Gly Asn Ser Thr Ala Gln Tyr Met Leu 145 150 155 160 Gly Phe Met Tyr Ala Thr Gly Val Gly Gly Ala Val Glu Arg Asp Gln 165 170 175 Ala Lys Ala Leu Leu Tyr His Thr Phe Ala Ala Glu Ala Gly Asn Thr 180 185 190 Lys Ser Glu Met Thr Leu Ala Tyr Arg Tyr His Ala Gly Ile Gly Ala 195 200 205 Pro Arg Asp Cys Asp Gln Ala Thr Tyr Tyr Tyr Lys Lys Val Ala Asp 210 215 220 Lys Ala Ile Glu Tyr Phe Arg Ser Gly Pro Pro Gly Gly His Asn Met 225 230 235 240 Ile Arg Glu Ser Tyr Arg Trp Ala Asp Glu Glu Gly Gly Val Tyr Gly 245 250 255 Glu Gly Ala Ser Val Ser Thr Ala Val Arg Asp Gly Thr His Ser Ser 260 265 270 Thr Glu Ala Ser Leu Glu Asp Val Leu Glu Tyr Leu Asp Leu Met Ser 275 280 285 Arg Lys Gly Glu Leu Lys Ala Thr Phe Ser Leu Gly Lys Met His Tyr 290 295 300 Glu Gly Gly Arg Gly Leu Pro Arg Asn Phe Arg Lys Ser Met Asn Tyr 305 310 315 320 Phe Arg Gln Val Ala Lys Arg Tyr Trp Asn Lys Asp Gly Ser Val Asn 325 330 335 Pro Asn His Pro Val Gly Val Glu Lys Leu Ala Ser Lys Ala Ala Gly 340 345 350 His Ile Gly Met Met Tyr Leu Arg Gly Glu Gly Val Glu Gln Asn Phe 355 360 365 Ala Thr Ala Gln Thr Trp Phe Arg Arg Gly Leu Ala Asn Gly Asp Ala 370 375 380 Leu Cys Gln His Glu Leu Gly Leu Met Tyr Leu His Gly Tyr Gly Val 385 390 395 400 Thr Pro Asp Ala Phe Arg Ala Ala Ser Gln Phe Lys Ala Ala Ala Glu 405 410 415 Gln Asp Phe Pro Ala Ala Glu Thr Arg Leu Gly Ala Leu Phe Leu Asp 420 425 430 Gln Gly Asp Val Gln Thr Ala Thr Arg Tyr Phe Glu Leu Ala Ala Arg 435 440 445 Trp Gly Trp Met Glu Ala Phe Tyr Tyr Leu Ala Glu Leu Ser Asn Asn 450 455 460 Gly Val Gly Arg Lys Arg His Cys Gly Met Ala Ala Ser Tyr Tyr Lys 465 470 475 480 Met Val Ala Glu Arg Ala Glu Val Ile His Ser Ser Phe Glu Glu Ala 485 490 495 Asn Thr Ala Tyr Glu Asn Gly Asp Lys Glu Arg Ala Leu Ile Pro Ala 500 505 510 Leu Met Ala Ala Glu Gln Gly Tyr Glu His Ala Gln Ser Asn Val Ala 515 520 525 Phe Leu Leu Asp Glu Gln Arg Ser Leu Phe Ala Ile Asp Thr Ile Leu 530 535 540 Pro Gly Ala Lys Lys Ser Arg Pro Ala Leu Leu Arg Asn Ala Ala Leu 545 550 555 560 Ala Leu Ile Tyr Trp Thr Arg Ser Ala Lys Gln Ala Asn Ile Asp Ser 565 570 575 Leu Leu Lys Met Gly Asp Tyr Tyr Leu Ala Gly Met Gly Ile Ala Ala 580 585 590 Asp Ala Glu Lys Ala Ser Thr Cys Tyr His Thr Ala Ala Glu Val His 595 600 605 Tyr Ser Ala Gln Ala Tyr Trp Asn Leu Gly Trp Met His Glu Asn Gly 610 615 620 Val Ala Val Asp Gln Asp Phe His Met Ala Lys Arg Tyr Tyr Asp Leu 625 630 635 640 Ala Leu Glu Thr Ser Ser Glu Ala Tyr Leu Pro Val Lys Leu Ser Leu 645 650 655 Leu Lys Leu Arg Met Arg Gly Tyr Trp Asn Trp Leu Thr Asn Gly Asp 660 665 670 Ile Asn Pro Ile Arg Glu Glu Glu Gly Lys Glu Pro Gln His Pro Phe 675 680 685 Arg 462326DNAAspergillus niger 46ctgtctggtt catgcacatc acttactagt tagcacttgt tcatattgtc attgtgtcaa 60ttttactcga ctgatcccaa ctttatcatt ctcctctgct ttacgccatt gtttctcttg 120cagtgagcat tgcgacttcg attatgcgcc cgtgattgtc tacgcccaac atccctccgc 180aaggacagaa caatcccagg atgaccgcgt cttcagaaac ccagtccgct ctggcggaat 240ctccacagag catagtatta catgtgctat gtccatcttt acctcctcct aaccgattca 300ctctccacga catctctcca tccaccacca tctccactct caaagctcga atcgcccaga 360ccattccgag tgaaccatcc cctgaaactc agaggctgat ataccggggg aagcccctta 420cgaacgacgc cgtggcgcta aatgatgtgt tagagtcatc aaatgtgcgt atgatcttat 480tcctatgccc ccttgtagtc ggcagatact aacagtgata cttgacccag gataccgagt 540attccatcca cctcgtcctt cctcctgccc cagttcccca tgcttcaact tctgccagag 600ctcctgctcc aatgccgcgg ggagcttcag gtaaccccgc gcctcagagt ccattctcat 660cgaaccgatt cacgccgcaa cacctgcctc acggacaaga gatcaggtat cgaggtcccg 720cattgcccgc tgttccccat gaggcagaga tcggactcgc cttgaggcgg aatatcgagg 780ctattcgtcg acagattgat atgcaggaac ggggcgggcc gttgggaggc gtcgcggcag 840gcaccgcggg ggctacttcc cactcgacca catccacgac tacgacagct tcgttcgctc 900agcaacctgc atggccacac gtaacacctg gtctatcgca cccaggacat tcgtccatct 960catctgattt cacaatggct tccggctcat cgggaacagc caatgtccat agcaacctcc 1020ccgaagaagt ccgattgcgc ctacaaatac tcagaaatca gattgcgttt ggcgaagagc 1080aactgaaccg gggggttgcg cccccaatgg accatataat tcgcatacgt acacaattgt 1140tcgctttgct cgacgaccaa tatcagaacc cacatgctga gcgcgacggc tcgattgaat 1200ctttgctcac tcgcgtcttt aatatctata cccgcgctga tcagctccgc gtctcacaag 1260ctagaaccat gcccacacct gttctatctg gacctcccaa tcccgcacca gggcaggctc 1320ctctgtacct tctctcgtca ccaaacggct atcaagccct ggttgcatct ccccgcggtg 1380cagaaacgat gcagtcttct ctcgatacac tccgagccat gcactccccg accggagcct 1440ctgcgcctcg tacaggtgct ccaccggaga ttcacaatgc gaacgcagtt gtcatggaga 1500acattgtccg acaggccgta ctcaaccaac gcatcgaaaa caacgggcaa ttgagcttca 1560cacgcaatct ccgacgcatg tggctatttg tgcgcttgta tttcttctgc tacatgttca 1620gcgaaccggg cacatggtct cgcgtggtgt atgtaaccct agccgtcctt gtctcactcc 1680tgtcagaaac cgggatccca cagcagttgt accgaatgct tgtggcgcca gtgcaacgac 1740acctggaagg actggttcat ttcgctccgg acgaaccgac tccagcacca cctggcacgc 1800agtcaactgg gcaagggaac gttcccactg ctcagccaac cggaatgcga caccagctgc 1860gccgcgtaga acggtccttg gcgcttttca ttgcaagctt ggttccgggc gtgggcgaga 1920gacacgtgga agtgcgcaat gccgcagaag cggcccggaa cgcagagcgt gcaagagagg 1980aagaagagcg acgtcgacag gaggaagagg ccaccaacgc aggcacgact ggtgaggctc 2040aggcacaggc acaggagagc agcgagaacg aacagagaga aacgggtgag aatgcaccca 2100acacgatacc ccaaactgag aattagccat agcggagcta ttagacttgt actgagtata 2160ctcggtgatt tgaaacttgt gttcatgatg tattgatagc tgcgatcata tttccatacc 2220gcttagtgcg gcaagatatc ttagtctatt agcgcaacat atatctactg cagtgtacct 2280tcaagtaatc tatccacatc cacccacaca taatccactc tatcta 2326471860DNAAspergillus nigerCDS(1)..(1860) 47atg acc gcg tct tca gaa acc cag tcc gct ctg gcg gaa tct cca cag 48Met Thr Ala Ser Ser Glu Thr Gln Ser Ala Leu Ala Glu Ser Pro Gln 1 5 10 15 agc ata gta tta cat gtg cta tgt cca tct tta cct cct cct aac cga 96Ser Ile Val Leu His Val Leu Cys Pro Ser Leu Pro Pro Pro Asn Arg 20 25 30 ttc act ctc cac gac atc tct cca tcc acc acc atc tcc act ctc aaa 144Phe Thr Leu His Asp Ile Ser Pro Ser Thr Thr Ile Ser Thr Leu Lys 35 40 45 gct cga atc gcc cag acc att ccg agt gaa cca tcc cct gaa act cag 192Ala Arg Ile Ala Gln Thr Ile Pro Ser Glu Pro Ser Pro Glu Thr Gln 50 55 60 agg ctg ata tac cgg ggg aag ccc ctt acg aac gac gcc gtg gcg cta 240Arg Leu Ile Tyr Arg Gly Lys Pro Leu Thr Asn Asp Ala Val Ala Leu 65 70 75 80 aat gat gtg tta gag tca tca aat gat acc gag tat tcc atc cac ctc 288Asn Asp Val Leu Glu Ser Ser Asn Asp Thr Glu Tyr Ser Ile His Leu 85 90 95 gtc ctt cct cct gcc cca gtt ccc cat gct tca act tct gcc aga gct 336Val Leu Pro Pro Ala Pro Val Pro His Ala Ser Thr Ser Ala Arg Ala 100 105 110 cct gct cca atg ccg cgg gga gct tca ggt aac ccc gcg cct cag agt 384Pro Ala Pro Met Pro Arg Gly Ala Ser Gly Asn Pro Ala Pro Gln Ser 115 120 125 cca ttc tca tcg aac cga ttc acg ccg caa cac ctg cct cac gga caa 432Pro Phe Ser Ser Asn Arg Phe Thr Pro Gln His Leu Pro His Gly Gln 130 135 140 gag atc agg tat cga ggt ccc gca ttg ccc gct gtt ccc cat gag gca 480Glu Ile Arg Tyr Arg Gly Pro Ala Leu Pro Ala Val Pro His Glu Ala 145 150 155 160 gag atc gga ctc gcc ttg agg cgg aat atc gag gct att cgt cga cag 528Glu Ile Gly Leu Ala Leu Arg Arg Asn Ile Glu Ala Ile Arg Arg Gln 165 170 175 att gat atg cag gaa cgg ggc ggg ccg ttg gga ggc gtc gcg gca ggc 576Ile Asp Met Gln Glu Arg Gly Gly Pro Leu Gly Gly Val Ala Ala Gly 180 185 190 acc gcg ggg gct act tcc cac tcg acc aca tcc acg act acg aca gct 624Thr Ala Gly Ala Thr Ser His Ser Thr Thr Ser Thr Thr Thr Thr Ala 195 200 205 tcg ttc gct cag caa cct gca tgg cca cac gta aca cct ggt cta tcg 672Ser Phe Ala Gln Gln Pro Ala Trp Pro His Val Thr Pro Gly Leu Ser 210 215 220 cac cca gga cat tcg tcc atc tca tct gat ttc aca atg gct tcc ggc 720His Pro Gly His Ser Ser Ile Ser Ser Asp Phe Thr Met Ala Ser Gly 225 230 235 240 tca tcg gga aca gcc aat gtc cat agc aac ctc ccc gaa gaa gtc cga 768Ser Ser Gly Thr Ala Asn Val His Ser Asn Leu Pro Glu Glu Val Arg 245 250 255 ttg cgc cta caa ata ctc aga aat cag att gcg ttt ggc gaa gag caa 816Leu Arg Leu Gln Ile Leu Arg Asn Gln Ile Ala Phe Gly Glu Glu Gln 260 265 270 ctg aac cgg ggg gtt gcg ccc cca atg gac cat ata att cgc ata cgt 864Leu Asn Arg Gly Val Ala Pro Pro Met Asp His Ile Ile Arg Ile Arg 275 280 285 aca caa ttg ttc gct ttg ctc gac gac caa tat cag aac cca cat gct 912Thr Gln Leu Phe Ala Leu Leu Asp Asp Gln Tyr Gln Asn Pro His Ala 290 295 300 gag cgc gac ggc tcg att gaa tct ttg ctc act cgc gtc ttt aat atc 960Glu Arg Asp Gly Ser Ile Glu Ser Leu Leu Thr Arg Val Phe Asn Ile 305 310 315 320 tat acc cgc gct gat cag ctc cgc gtc tca caa gct aga acc atg ccc 1008Tyr Thr Arg Ala Asp Gln Leu Arg Val Ser Gln Ala Arg Thr Met Pro 325 330 335 aca cct gtt cta tct gga cct ccc aat ccc gca cca ggg cag gct cct 1056Thr Pro Val Leu Ser Gly Pro Pro Asn Pro Ala Pro Gly Gln Ala Pro 340 345 350 ctg tac ctt ctc tcg tca cca aac ggc tat caa gcc ctg gtt gca tct 1104Leu Tyr Leu Leu Ser Ser Pro Asn Gly Tyr Gln Ala Leu Val Ala Ser 355 360 365 ccc cgc ggt gca gaa acg atg cag tct tct ctc gat aca ctc cga gcc 1152Pro Arg Gly Ala Glu Thr Met Gln Ser Ser Leu Asp Thr Leu Arg Ala 370 375

380 atg cac tcc ccg acc gga gcc tct gcg cct cgt aca ggt gct cca ccg 1200Met His Ser Pro Thr Gly Ala Ser Ala Pro Arg Thr Gly Ala Pro Pro 385 390 395 400 gag att cac aat gcg aac gca gtt gtc atg gag aac att gtc cga cag 1248Glu Ile His Asn Ala Asn Ala Val Val Met Glu Asn Ile Val Arg Gln 405 410 415 gcc gta ctc aac caa cgc atc gaa aac aac ggg caa ttg agc ttc aca 1296Ala Val Leu Asn Gln Arg Ile Glu Asn Asn Gly Gln Leu Ser Phe Thr 420 425 430 cgc aat ctc cga cgc atg tgg cta ttt gtg cgc ttg tat ttc ttc tgc 1344Arg Asn Leu Arg Arg Met Trp Leu Phe Val Arg Leu Tyr Phe Phe Cys 435 440 445 tac atg ttc agc gaa ccg ggc aca tgg tct cgc gtg gtg tat gta acc 1392Tyr Met Phe Ser Glu Pro Gly Thr Trp Ser Arg Val Val Tyr Val Thr 450 455 460 cta gcc gtc ctt gtc tca ctc ctg tca gaa acc ggg atc cca cag cag 1440Leu Ala Val Leu Val Ser Leu Leu Ser Glu Thr Gly Ile Pro Gln Gln 465 470 475 480 ttg tac cga atg ctt gtg gcg cca gtg caa cga cac ctg gaa gga ctg 1488Leu Tyr Arg Met Leu Val Ala Pro Val Gln Arg His Leu Glu Gly Leu 485 490 495 gtt cat ttc gct ccg gac gaa ccg act cca gca cca cct ggc acg cag 1536Val His Phe Ala Pro Asp Glu Pro Thr Pro Ala Pro Pro Gly Thr Gln 500 505 510 tca act ggg caa ggg aac gtt ccc act gct cag cca acc gga atg cga 1584Ser Thr Gly Gln Gly Asn Val Pro Thr Ala Gln Pro Thr Gly Met Arg 515 520 525 cac cag ctg cgc cgc gta gaa cgg tcc ttg gcg ctt ttc att gca agc 1632His Gln Leu Arg Arg Val Glu Arg Ser Leu Ala Leu Phe Ile Ala Ser 530 535 540 ttg gtt ccg ggc gtg ggc gag aga cac gtg gaa gtg cgc aat gcc gca 1680Leu Val Pro Gly Val Gly Glu Arg His Val Glu Val Arg Asn Ala Ala 545 550 555 560 gaa gcg gcc cgg aac gca gag cgt gca aga gag gaa gaa gag cga cgt 1728Glu Ala Ala Arg Asn Ala Glu Arg Ala Arg Glu Glu Glu Glu Arg Arg 565 570 575 cga cag gag gaa gag gcc acc aac gca ggc acg act ggt gag gct cag 1776Arg Gln Glu Glu Glu Ala Thr Asn Ala Gly Thr Thr Gly Glu Ala Gln 580 585 590 gca cag gca cag gag agc agc gag aac gaa cag aga gaa acg ggt gag 1824Ala Gln Ala Gln Glu Ser Ser Glu Asn Glu Gln Arg Glu Thr Gly Glu 595 600 605 aat gca ccc aac acg ata ccc caa act gag aat tag 1860Asn Ala Pro Asn Thr Ile Pro Gln Thr Glu Asn 610 615 48619PRTAspergillus niger 48Met Thr Ala Ser Ser Glu Thr Gln Ser Ala Leu Ala Glu Ser Pro Gln 1 5 10 15 Ser Ile Val Leu His Val Leu Cys Pro Ser Leu Pro Pro Pro Asn Arg 20 25 30 Phe Thr Leu His Asp Ile Ser Pro Ser Thr Thr Ile Ser Thr Leu Lys 35 40 45 Ala Arg Ile Ala Gln Thr Ile Pro Ser Glu Pro Ser Pro Glu Thr Gln 50 55 60 Arg Leu Ile Tyr Arg Gly Lys Pro Leu Thr Asn Asp Ala Val Ala Leu 65 70 75 80 Asn Asp Val Leu Glu Ser Ser Asn Asp Thr Glu Tyr Ser Ile His Leu 85 90 95 Val Leu Pro Pro Ala Pro Val Pro His Ala Ser Thr Ser Ala Arg Ala 100 105 110 Pro Ala Pro Met Pro Arg Gly Ala Ser Gly Asn Pro Ala Pro Gln Ser 115 120 125 Pro Phe Ser Ser Asn Arg Phe Thr Pro Gln His Leu Pro His Gly Gln 130 135 140 Glu Ile Arg Tyr Arg Gly Pro Ala Leu Pro Ala Val Pro His Glu Ala 145 150 155 160 Glu Ile Gly Leu Ala Leu Arg Arg Asn Ile Glu Ala Ile Arg Arg Gln 165 170 175 Ile Asp Met Gln Glu Arg Gly Gly Pro Leu Gly Gly Val Ala Ala Gly 180 185 190 Thr Ala Gly Ala Thr Ser His Ser Thr Thr Ser Thr Thr Thr Thr Ala 195 200 205 Ser Phe Ala Gln Gln Pro Ala Trp Pro His Val Thr Pro Gly Leu Ser 210 215 220 His Pro Gly His Ser Ser Ile Ser Ser Asp Phe Thr Met Ala Ser Gly 225 230 235 240 Ser Ser Gly Thr Ala Asn Val His Ser Asn Leu Pro Glu Glu Val Arg 245 250 255 Leu Arg Leu Gln Ile Leu Arg Asn Gln Ile Ala Phe Gly Glu Glu Gln 260 265 270 Leu Asn Arg Gly Val Ala Pro Pro Met Asp His Ile Ile Arg Ile Arg 275 280 285 Thr Gln Leu Phe Ala Leu Leu Asp Asp Gln Tyr Gln Asn Pro His Ala 290 295 300 Glu Arg Asp Gly Ser Ile Glu Ser Leu Leu Thr Arg Val Phe Asn Ile 305 310 315 320 Tyr Thr Arg Ala Asp Gln Leu Arg Val Ser Gln Ala Arg Thr Met Pro 325 330 335 Thr Pro Val Leu Ser Gly Pro Pro Asn Pro Ala Pro Gly Gln Ala Pro 340 345 350 Leu Tyr Leu Leu Ser Ser Pro Asn Gly Tyr Gln Ala Leu Val Ala Ser 355 360 365 Pro Arg Gly Ala Glu Thr Met Gln Ser Ser Leu Asp Thr Leu Arg Ala 370 375 380 Met His Ser Pro Thr Gly Ala Ser Ala Pro Arg Thr Gly Ala Pro Pro 385 390 395 400 Glu Ile His Asn Ala Asn Ala Val Val Met Glu Asn Ile Val Arg Gln 405 410 415 Ala Val Leu Asn Gln Arg Ile Glu Asn Asn Gly Gln Leu Ser Phe Thr 420 425 430 Arg Asn Leu Arg Arg Met Trp Leu Phe Val Arg Leu Tyr Phe Phe Cys 435 440 445 Tyr Met Phe Ser Glu Pro Gly Thr Trp Ser Arg Val Val Tyr Val Thr 450 455 460 Leu Ala Val Leu Val Ser Leu Leu Ser Glu Thr Gly Ile Pro Gln Gln 465 470 475 480 Leu Tyr Arg Met Leu Val Ala Pro Val Gln Arg His Leu Glu Gly Leu 485 490 495 Val His Phe Ala Pro Asp Glu Pro Thr Pro Ala Pro Pro Gly Thr Gln 500 505 510 Ser Thr Gly Gln Gly Asn Val Pro Thr Ala Gln Pro Thr Gly Met Arg 515 520 525 His Gln Leu Arg Arg Val Glu Arg Ser Leu Ala Leu Phe Ile Ala Ser 530 535 540 Leu Val Pro Gly Val Gly Glu Arg His Val Glu Val Arg Asn Ala Ala 545 550 555 560 Glu Ala Ala Arg Asn Ala Glu Arg Ala Arg Glu Glu Glu Glu Arg Arg 565 570 575 Arg Gln Glu Glu Glu Ala Thr Asn Ala Gly Thr Thr Gly Glu Ala Gln 580 585 590 Ala Gln Ala Gln Glu Ser Ser Glu Asn Glu Gln Arg Glu Thr Gly Glu 595 600 605 Asn Ala Pro Asn Thr Ile Pro Gln Thr Glu Asn 610 615 492941DNAAspergillus niger 49tcaggtactc caggtgaact agttggtact actaagcagg aactcccccc ctctctctct 60tcccaaaggt gccttgaaac cctcccggcc atctttggtg cctccatctc atccttttgg 120ctcttgccag ttccccctct tcatctctaa ccttccctac tgggatctat atttccctac 180ttctctccac tggtctatgt atgcctgagt tcaagatctc cgcttctttg gagggccacg 240gcgatgatgt aagaacatcc cggctgttga cgctggccac ctttcacctt caacccccct 300ccattgtcgc ccactgacgc ctctttctct cactctcagg ttcgcgccgt ggcctttccg 360aatcccaatg ctatattttc ggcgtcgcga gatgcaacag tccgactctg gaaactagtc 420tctaccccac ctccggcata tgactacacc atcacctctc acggccaggc cttcatcaac 480gctttggcat actacccacc taccccccag tttcccgatg gacttgtcct ttccggtggt 540caagatacta tcattgaagc cagacaacca ggcaaagctg ccgacgataa cgcggatgct 600atgctcttgg gccatacaca taatgtctgt gcgctggatg tgtcacatga tggcggatgg 660gtagtcagcg gaagctggga ctcgacagct agactatgga aagtgggtaa atgggaaacc 720gatgtcgtgc tggagggcca tcaaggaagt gtttggacgg tgcttgccta tgacaaggat 780acggtcatca caggtaggcg cgccttaccc ctgtatatga gacagatggg tcgttctatg 840gatatgctaa taattcccca ggctgcgcgg acaaaataat acgtattttt aacacctctg 900gcacactgct gagaacaatc gaaaattcac aggacgttgt gagagctctt tgcaaggttc 960ccgcttcgaa ccccaccggg gcgcactttg cttcggcgag caacgatgga gtgattcgtc 1020tttttaccat acaaggccaa ctcgtcgggg agatgcatgg ccacgagagc ttcatttatt 1080ctctggccgc tttgccttcg ggtgagttag tcagttccgg agaagatcgg acggtgagag 1140tctgggatgg tacgcagtgc gtacagacga tcacacaccc tgcgatctct gtctggagcg 1200tcgcagtatg caaggagacc ggcgacattg ttacaggagc cagtgaccga atcacacgcg 1260tgtttagcag gagccaggag cgcgtggcaa gcccagaagt agtacaacag ttcgagaaga 1320ctgtgaagga gtcggcaatc ccagagcagc agattgggaa gatcaacaaa gataagcttc 1380cgggtacgga gtttctcagg cagaaatccg ggaccaagga cgggcaggtg cagatgatcc 1440gtgaggccga tggtagcgtt actgctcaca cttggtcagc ggcctcacgg gaatgggttg 1500cggttggcac ggtagttgat tccgctgcca gcagtggaag gaaaacggag tatctgggtc 1560aagactacga ctatgtcttt gatgtcgacg tggaagacgg caaacccccc ctcaaattgc 1620catacaacgt ctctcaaaac ccctacgagg ctgcgaccaa gtttatccag gacaacgaac 1680tgtcgatgaa ttaccttgat caagttgctc agttcatcgt tcagaatacg caaggtgcga 1740ctcttggtca gacgtctcag gggccgacgc ccgcgggggc cgatccctgg ggtcaagaga 1800ggcgttatcg tcctgaagat gcgcagtcgc cccctgctcc tgaggcccga ccgaaggtcc 1860ttccgcaaaa aacatatctt tccataaaat ctgctaatct taaactgatc gctaagaagt 1920tgcaagagct gaaccaacac gtcatatcct ccggatcgaa agacctgtcg ctcagccctt 1980cagagttgga gacggtggca accttgtgtg gtcagttgga gtcttcgaat gttgagcagt 2040ctccggcagt ggaggctggt gttgttttac tatacaaggt cgcaaccgtc tggcccgtcg 2100caagcagact accaggtctt gatcttctcc gcttgtccgc cgctgctact cccgtgactg 2160ccactgcaga ttacgatggc aaggatctca tctcagggat taagtctagc ggggtgttcg 2220attcaccgtt caatgtcaat aatgcgatgc tgtcaatacg catgctcgcc aaccttttcg 2280aaacggatgc gggacgtgac ctggccacta gcaagtttga gcagattctg agcggcgtca 2340agtccgcttt aaccaacagt gggacgacgc cgaaccgaaa tctcaccatt gccattacaa 2400cactctacat caactttgcc gtttacctga cctctgcggg cagagaatcg atgcctgagt 2460catcggaaca ggctctggtg cttctcggcg agctaacgac attgattacc ggtgaaaagg 2520actctgaagc agtctaccgc ggccttgtgg ctctagggac cttgatcaag ggactagggg 2580aagaagtcag gactgcggcc aaggaagtgt acgatgtcga tgatgttttg aagaaggttt 2640caagctctgg tcttggtaaa gaaccaagaa tcaagggtat cataggcgag atcaaggagt 2700cgttatcatc aaggtataaa atgttgaggc ccgggtctta agtcaactca tacctcaagg 2760agcgtcttgt gttctgcttc accactgctg tctatagcac gtatacatct ctagttcacg 2820atagcgatac acactacact gcatgaattc aacattggac atattccaaa tcagtgctaa 2880cactaggtat cgtatagtgc tatacattca atgtaacggc aacgtaaata atcactttga 2940c 2941502331DNAAspergillus nigerCDS(1)..(2331) 50atg cct gag ttc aag atc tcc gct tct ttg gag ggc cac ggc gat gat 48Met Pro Glu Phe Lys Ile Ser Ala Ser Leu Glu Gly His Gly Asp Asp 1 5 10 15 gtt cgc gcc gtg gcc ttt ccg aat ccc aat gct ata ttt tcg gcg tcg 96Val Arg Ala Val Ala Phe Pro Asn Pro Asn Ala Ile Phe Ser Ala Ser 20 25 30 cga gat gca aca gtc cga ctc tgg aaa cta gtc tct acc cca cct ccg 144Arg Asp Ala Thr Val Arg Leu Trp Lys Leu Val Ser Thr Pro Pro Pro 35 40 45 gca tat gac tac acc atc acc tct cac ggc cag gcc ttc atc aac gct 192Ala Tyr Asp Tyr Thr Ile Thr Ser His Gly Gln Ala Phe Ile Asn Ala 50 55 60 ttg gca tac tac cca cct acc ccc cag ttt ccc gat gga ctt gtc ctt 240Leu Ala Tyr Tyr Pro Pro Thr Pro Gln Phe Pro Asp Gly Leu Val Leu 65 70 75 80 tcc ggt ggt caa gat act atc att gaa gcc aga caa cca ggc aaa gct 288Ser Gly Gly Gln Asp Thr Ile Ile Glu Ala Arg Gln Pro Gly Lys Ala 85 90 95 gcc gac gat aac gcg gat gct atg ctc ttg ggc cat aca cat aat gtc 336Ala Asp Asp Asn Ala Asp Ala Met Leu Leu Gly His Thr His Asn Val 100 105 110 tgt gcg ctg gat gtg tca cat gat ggc gga tgg gta gtc agc gga agc 384Cys Ala Leu Asp Val Ser His Asp Gly Gly Trp Val Val Ser Gly Ser 115 120 125 tgg gac tcg aca gct aga cta tgg aaa gtg ggt aaa tgg gaa acc gat 432Trp Asp Ser Thr Ala Arg Leu Trp Lys Val Gly Lys Trp Glu Thr Asp 130 135 140 gtc gtg ctg gag ggc cat caa gga agt gtt tgg acg gtg ctt gcc tat 480Val Val Leu Glu Gly His Gln Gly Ser Val Trp Thr Val Leu Ala Tyr 145 150 155 160 gac aag gat acg gtc atc aca ggc tgc gcg gac aaa ata ata cgt att 528Asp Lys Asp Thr Val Ile Thr Gly Cys Ala Asp Lys Ile Ile Arg Ile 165 170 175 ttt aac acc tct ggc aca ctg ctg aga aca atc gaa aat tca cag gac 576Phe Asn Thr Ser Gly Thr Leu Leu Arg Thr Ile Glu Asn Ser Gln Asp 180 185 190 gtt gtg aga gct ctt tgc aag gtt ccc gct tcg aac ccc acc ggg gcg 624Val Val Arg Ala Leu Cys Lys Val Pro Ala Ser Asn Pro Thr Gly Ala 195 200 205 cac ttt gct tcg gcg agc aac gat gga gtg att cgt ctt ttt acc ata 672His Phe Ala Ser Ala Ser Asn Asp Gly Val Ile Arg Leu Phe Thr Ile 210 215 220 caa ggc caa ctc gtc ggg gag atg cat ggc cac gag agc ttc att tat 720Gln Gly Gln Leu Val Gly Glu Met His Gly His Glu Ser Phe Ile Tyr 225 230 235 240 tct ctg gcc gct ttg cct tcg ggt gag tta gtc agt tcc gga gaa gat 768Ser Leu Ala Ala Leu Pro Ser Gly Glu Leu Val Ser Ser Gly Glu Asp 245 250 255 cgg acg gtg aga gtc tgg gat ggt acg cag tgc gta cag acg atc aca 816Arg Thr Val Arg Val Trp Asp Gly Thr Gln Cys Val Gln Thr Ile Thr 260 265 270 cac cct gcg atc tct gtc tgg agc gtc gca gta tgc aag gag acc ggc 864His Pro Ala Ile Ser Val Trp Ser Val Ala Val Cys Lys Glu Thr Gly 275 280 285 gac att gtt aca gga gcc agt gac cga atc aca cgc gtg ttt agc agg 912Asp Ile Val Thr Gly Ala Ser Asp Arg Ile Thr Arg Val Phe Ser Arg 290 295 300 agc cag gag cgc gtg gca agc cca gaa gta gta caa cag ttc gag aag 960Ser Gln Glu Arg Val Ala Ser Pro Glu Val Val Gln Gln Phe Glu Lys 305 310 315 320 act gtg aag gag tcg gca atc cca gag cag cag att ggg aag atc aac 1008Thr Val Lys Glu Ser Ala Ile Pro Glu Gln Gln Ile Gly Lys Ile Asn 325 330 335 aaa gat aag ctt ccg ggt acg gag ttt ctc agg cag aaa tcc ggg acc 1056Lys Asp Lys Leu Pro Gly Thr Glu Phe Leu Arg Gln Lys Ser Gly Thr 340 345 350 aag gac ggg cag gtg cag atg atc cgt gag gcc gat ggt agc gtt act 1104Lys Asp Gly Gln Val Gln Met Ile Arg Glu Ala Asp Gly Ser Val Thr 355 360 365 gct cac act tgg tca gcg gcc tca cgg gaa tgg gtt gcg gtt ggc acg 1152Ala His Thr Trp Ser Ala Ala Ser Arg Glu Trp Val Ala Val Gly Thr 370 375 380 gta gtt gat tcc gct gcc agc agt gga agg aaa acg gag tat ctg ggt 1200Val Val Asp Ser Ala Ala Ser Ser Gly Arg Lys Thr Glu Tyr Leu Gly 385 390 395 400 caa gac tac gac tat gtc ttt gat gtc gac gtg gaa gac ggc aaa ccc 1248Gln Asp Tyr Asp Tyr Val Phe Asp Val Asp Val Glu Asp Gly Lys Pro 405 410 415 ccc ctc aaa ttg cca tac aac gtc tct caa aac ccc tac gag gct gcg 1296Pro Leu Lys Leu Pro Tyr Asn Val Ser Gln Asn Pro Tyr Glu Ala Ala 420 425 430 acc aag ttt atc cag gac aac gaa ctg tcg atg aat tac ctt gat caa 1344Thr Lys Phe Ile Gln Asp Asn Glu Leu Ser Met Asn Tyr Leu Asp Gln 435 440 445 gtt gct cag ttc atc gtt cag aat acg caa ggt gcg act ctt gag agg 1392Val Ala Gln Phe Ile Val Gln Asn Thr Gln Gly Ala Thr Leu Glu Arg 450 455 460 cgt tat cgt cct gaa gat gcg cag tcg ccc cct gct cct gag gcc cga 1440Arg Tyr Arg Pro Glu Asp Ala Gln Ser Pro Pro Ala Pro Glu Ala Arg 465 470 475 480 ccg aag gtc ctt ccg caa aaa aca tat ctt tcc ata aaa tct gct aat 1488Pro Lys Val Leu Pro Gln Lys Thr Tyr Leu Ser Ile Lys Ser Ala Asn 485 490 495 ctt aaa ctg atc gct aag aag ttg caa gag ctg aac caa cac gtc ata 1536Leu Lys Leu Ile Ala Lys Lys Leu Gln Glu Leu Asn Gln His Val Ile 500 505 510 tcc tcc gga tcg aaa gac ctg tcg ctc agc cct tca gag ttg gag acg 1584Ser Ser Gly Ser Lys Asp Leu Ser Leu Ser Pro Ser Glu Leu Glu Thr 515 520 525 gtg

gca acc ttg tgt ggt cag ttg gag tct tcg aat gtt gag cag tct 1632Val Ala Thr Leu Cys Gly Gln Leu Glu Ser Ser Asn Val Glu Gln Ser 530 535 540 ccg gca gtg gag gct ggt gtt gtt tta cta tac aag gtc gca acc gtc 1680Pro Ala Val Glu Ala Gly Val Val Leu Leu Tyr Lys Val Ala Thr Val 545 550 555 560 tgg ccc gtc gca agc aga cta cca ggt ctt gat ctt ctc cgc ttg tcc 1728Trp Pro Val Ala Ser Arg Leu Pro Gly Leu Asp Leu Leu Arg Leu Ser 565 570 575 gcc gct gct act ccc gtg act gcc act gca gat tac gat ggc aag gat 1776Ala Ala Ala Thr Pro Val Thr Ala Thr Ala Asp Tyr Asp Gly Lys Asp 580 585 590 ctc atc tca ggg att aag tct agc ggg gtg ttc gat tca ccg ttc aat 1824Leu Ile Ser Gly Ile Lys Ser Ser Gly Val Phe Asp Ser Pro Phe Asn 595 600 605 gtc aat aat gcg atg ctg tca ata cgc atg ctc gcc aac ctt ttc gaa 1872Val Asn Asn Ala Met Leu Ser Ile Arg Met Leu Ala Asn Leu Phe Glu 610 615 620 acg gat gcg gga cgt gac ctg gcc act agc aag ttt gag cag att ctg 1920Thr Asp Ala Gly Arg Asp Leu Ala Thr Ser Lys Phe Glu Gln Ile Leu 625 630 635 640 agc ggc gtc aag tcc gct tta acc aac agt ggg acg acg ccg aac cga 1968Ser Gly Val Lys Ser Ala Leu Thr Asn Ser Gly Thr Thr Pro Asn Arg 645 650 655 aat ctc acc att gcc att aca aca ctc tac atc aac ttt gcc gtt tac 2016Asn Leu Thr Ile Ala Ile Thr Thr Leu Tyr Ile Asn Phe Ala Val Tyr 660 665 670 ctg acc tct gcg ggc aga gaa tcg atg cct gag tca tcg gaa cag gct 2064Leu Thr Ser Ala Gly Arg Glu Ser Met Pro Glu Ser Ser Glu Gln Ala 675 680 685 ctg gtg ctt ctc ggc gag cta acg aca ttg att acc ggt gaa aag gac 2112Leu Val Leu Leu Gly Glu Leu Thr Thr Leu Ile Thr Gly Glu Lys Asp 690 695 700 tct gaa gca gtc tac cgc ggc ctt gtg gct cta ggg acc ttg atc aag 2160Ser Glu Ala Val Tyr Arg Gly Leu Val Ala Leu Gly Thr Leu Ile Lys 705 710 715 720 gga cta ggg gaa gaa gtc agg act gcg gcc aag gaa gtg tac gat gtc 2208Gly Leu Gly Glu Glu Val Arg Thr Ala Ala Lys Glu Val Tyr Asp Val 725 730 735 gat gat gtt ttg aag aag gtt tca agc tct ggt ctt ggt aaa gaa cca 2256Asp Asp Val Leu Lys Lys Val Ser Ser Ser Gly Leu Gly Lys Glu Pro 740 745 750 aga atc aag ggt atc ata ggc gag atc aag gag tcg tta tca tca agg 2304Arg Ile Lys Gly Ile Ile Gly Glu Ile Lys Glu Ser Leu Ser Ser Arg 755 760 765 tat aaa atg ttg agg ccc ggg tct taa 2331Tyr Lys Met Leu Arg Pro Gly Ser 770 775 51776PRTAspergillus niger 51Met Pro Glu Phe Lys Ile Ser Ala Ser Leu Glu Gly His Gly Asp Asp 1 5 10 15 Val Arg Ala Val Ala Phe Pro Asn Pro Asn Ala Ile Phe Ser Ala Ser 20 25 30 Arg Asp Ala Thr Val Arg Leu Trp Lys Leu Val Ser Thr Pro Pro Pro 35 40 45 Ala Tyr Asp Tyr Thr Ile Thr Ser His Gly Gln Ala Phe Ile Asn Ala 50 55 60 Leu Ala Tyr Tyr Pro Pro Thr Pro Gln Phe Pro Asp Gly Leu Val Leu 65 70 75 80 Ser Gly Gly Gln Asp Thr Ile Ile Glu Ala Arg Gln Pro Gly Lys Ala 85 90 95 Ala Asp Asp Asn Ala Asp Ala Met Leu Leu Gly His Thr His Asn Val 100 105 110 Cys Ala Leu Asp Val Ser His Asp Gly Gly Trp Val Val Ser Gly Ser 115 120 125 Trp Asp Ser Thr Ala Arg Leu Trp Lys Val Gly Lys Trp Glu Thr Asp 130 135 140 Val Val Leu Glu Gly His Gln Gly Ser Val Trp Thr Val Leu Ala Tyr 145 150 155 160 Asp Lys Asp Thr Val Ile Thr Gly Cys Ala Asp Lys Ile Ile Arg Ile 165 170 175 Phe Asn Thr Ser Gly Thr Leu Leu Arg Thr Ile Glu Asn Ser Gln Asp 180 185 190 Val Val Arg Ala Leu Cys Lys Val Pro Ala Ser Asn Pro Thr Gly Ala 195 200 205 His Phe Ala Ser Ala Ser Asn Asp Gly Val Ile Arg Leu Phe Thr Ile 210 215 220 Gln Gly Gln Leu Val Gly Glu Met His Gly His Glu Ser Phe Ile Tyr 225 230 235 240 Ser Leu Ala Ala Leu Pro Ser Gly Glu Leu Val Ser Ser Gly Glu Asp 245 250 255 Arg Thr Val Arg Val Trp Asp Gly Thr Gln Cys Val Gln Thr Ile Thr 260 265 270 His Pro Ala Ile Ser Val Trp Ser Val Ala Val Cys Lys Glu Thr Gly 275 280 285 Asp Ile Val Thr Gly Ala Ser Asp Arg Ile Thr Arg Val Phe Ser Arg 290 295 300 Ser Gln Glu Arg Val Ala Ser Pro Glu Val Val Gln Gln Phe Glu Lys 305 310 315 320 Thr Val Lys Glu Ser Ala Ile Pro Glu Gln Gln Ile Gly Lys Ile Asn 325 330 335 Lys Asp Lys Leu Pro Gly Thr Glu Phe Leu Arg Gln Lys Ser Gly Thr 340 345 350 Lys Asp Gly Gln Val Gln Met Ile Arg Glu Ala Asp Gly Ser Val Thr 355 360 365 Ala His Thr Trp Ser Ala Ala Ser Arg Glu Trp Val Ala Val Gly Thr 370 375 380 Val Val Asp Ser Ala Ala Ser Ser Gly Arg Lys Thr Glu Tyr Leu Gly 385 390 395 400 Gln Asp Tyr Asp Tyr Val Phe Asp Val Asp Val Glu Asp Gly Lys Pro 405 410 415 Pro Leu Lys Leu Pro Tyr Asn Val Ser Gln Asn Pro Tyr Glu Ala Ala 420 425 430 Thr Lys Phe Ile Gln Asp Asn Glu Leu Ser Met Asn Tyr Leu Asp Gln 435 440 445 Val Ala Gln Phe Ile Val Gln Asn Thr Gln Gly Ala Thr Leu Glu Arg 450 455 460 Arg Tyr Arg Pro Glu Asp Ala Gln Ser Pro Pro Ala Pro Glu Ala Arg 465 470 475 480 Pro Lys Val Leu Pro Gln Lys Thr Tyr Leu Ser Ile Lys Ser Ala Asn 485 490 495 Leu Lys Leu Ile Ala Lys Lys Leu Gln Glu Leu Asn Gln His Val Ile 500 505 510 Ser Ser Gly Ser Lys Asp Leu Ser Leu Ser Pro Ser Glu Leu Glu Thr 515 520 525 Val Ala Thr Leu Cys Gly Gln Leu Glu Ser Ser Asn Val Glu Gln Ser 530 535 540 Pro Ala Val Glu Ala Gly Val Val Leu Leu Tyr Lys Val Ala Thr Val 545 550 555 560 Trp Pro Val Ala Ser Arg Leu Pro Gly Leu Asp Leu Leu Arg Leu Ser 565 570 575 Ala Ala Ala Thr Pro Val Thr Ala Thr Ala Asp Tyr Asp Gly Lys Asp 580 585 590 Leu Ile Ser Gly Ile Lys Ser Ser Gly Val Phe Asp Ser Pro Phe Asn 595 600 605 Val Asn Asn Ala Met Leu Ser Ile Arg Met Leu Ala Asn Leu Phe Glu 610 615 620 Thr Asp Ala Gly Arg Asp Leu Ala Thr Ser Lys Phe Glu Gln Ile Leu 625 630 635 640 Ser Gly Val Lys Ser Ala Leu Thr Asn Ser Gly Thr Thr Pro Asn Arg 645 650 655 Asn Leu Thr Ile Ala Ile Thr Thr Leu Tyr Ile Asn Phe Ala Val Tyr 660 665 670 Leu Thr Ser Ala Gly Arg Glu Ser Met Pro Glu Ser Ser Glu Gln Ala 675 680 685 Leu Val Leu Leu Gly Glu Leu Thr Thr Leu Ile Thr Gly Glu Lys Asp 690 695 700 Ser Glu Ala Val Tyr Arg Gly Leu Val Ala Leu Gly Thr Leu Ile Lys 705 710 715 720 Gly Leu Gly Glu Glu Val Arg Thr Ala Ala Lys Glu Val Tyr Asp Val 725 730 735 Asp Asp Val Leu Lys Lys Val Ser Ser Ser Gly Leu Gly Lys Glu Pro 740 745 750 Arg Ile Lys Gly Ile Ile Gly Glu Ile Lys Glu Ser Leu Ser Ser Arg 755 760 765 Tyr Lys Met Leu Arg Pro Gly Ser 770 775 522505DNAAspergillus niger 52acctaatctc ttcctttcac ctgatctgct gcctcttccc tccccatccc ctcccctcgt 60cttcccctcc caaaaagaat ccgccggttt ttccaccttt tccctttctt atccttttcc 120cctcccctcc tcggtggtgt gtatgtctgt aacgggcctt ttatcttcgt gaacagttta 180attgtgatcc actagaaatc atgggtcaga ccttgtccga gcccgtggtc gataaggtga 240gtgaaacctg aagcattgcg tttgcctgtc tggtctttga tcctcctccc ccacactgca 300cgcatcttac gccatttcaa tctgtccacc atccgcgttg ccattctgtg ccatacccgc 360ccatacttcc ctttttctat catttgtcca acatgatcac tgcccaaaag ttttatttgc 420tacagcgttc tttttcatcc ctcctcttgt gtgtcgcgcc tgcaatgtgg tgatcatccc 480ccctgcgggg ttcctccaca ggatgatgaa aatttcctgc acgcggtttc gggcggatga 540cctgtcacca ttccattgtg ttccggacct cttttcgatt tcctacatcc tcactaacca 600cattcggtgc tatctagact tccgctgaag gtcaagatga gtgctgtata tacggtgttt 660ccgccatgca aggatggcga atcagcatgg aggatgccca tgccgctgtc ctcgacctcc 720aggccaagta ctccgagcag gatgaaaagc cgaccgaccc cgataaacga ctcgctttct 780ttggtgtata tgacgggcac ggtggagaca aagtagcatt attcgccgga gaaaacgtcc 840acaagatcgt cgcgaagcag gactcctttg ccaaaggtga tatcgaacag gccctgaagg 900atggcttcct cgctaccgac cgggctattt tggaaggttc gtgagcgcta tgatcggagt 960tggggcaacc ttcccacccg cctctccctt tccttcctgt ttcctgtccc tccgaggtag 1020caacgaactg acccagctag acccgaaata tgaggaggaa gtgtctggct gcaccgcagc 1080cgtcagcgtt atctcgaagc acaagatctg ggtggtatgt attctagcca tggccgtttt 1140ggagacgacg ttgggtttgg ttcattcgtt gactggttct aggccaatgc tggtgattct 1200cgctctgtac tgggtgtcaa gggtcgcgca aagcctctgt catttgacca caagcctcag 1260aatgaaggta cacatacccc acatctcgat taccgaccag ttttgatgat gctgacatga 1320ccaaataaaa acaggcgaga aagcccgtat cagcgctgct ggtggtttcg ttgacttcgg 1380ccgtgtcaac ggcaacctgg ccttgtcgcg ggccattggt gacttcgagt tcaagaagag 1440ccccgagttg tctcctgagc agcagatcgt cactgcctat cccgacgtca ctgtgcacga 1500tctcagtgac gatgacgagt tcctcgtaat tgcctgtgac ggtgggtctt actctggtga 1560tgcgggggtg agggtcttga agatcgctaa cttttcgaaa ttgcaggtat ctgggattgt 1620cagtcctccc aatcagtggt ggaattcgtc cgccgtggga ttgccgcgaa gcaggatctg 1680tatcggatct gtgagaacat gatggacaac tgcctggcct ccaacagtga aaccggcggc 1740gttggctgcg acaatatgac aatggtcatc atcggtctcc tgaacggtag gaccaaagag 1800gagtggtaca accagatcgc tgagcgcgtg gcgaacggcg acggcccttg cgctccgccc 1860gagtacggca agtctctcga ggatcccacg gcctccaatt ccaatcccta ctgactgaac 1920ggggggtgca gctgagttcc gcggccccgg tatccggaat caattcgagg agaacccgga 1980tgactttgac atggaaaacg accgtgcgcg tggcttcagc gtccgctcgg gccgcatcat 2040cctcttgggg gacggcactg aattgattcc ggagcagaac gatgacgaac tctttgatca 2100ggctgaggaa gaccaggacc ttgtcaatca ggtgcaccgt gattcgcctg atgcggctcg 2160gaatgaacgg gagggaactc ctgggcctca gtctaaggat acttctcgaa cggacgccgc 2220tgagatatcg gagtcgccgt ctaccaccgc ggagggttcg tccggcagtg gccctggaac 2280gccgcaaaag cctacgagtt cgtagtcatg atggatctct tgcatttctc cttaatatgt 2340cttttctttt ttttcctcct tttttcccct tgccgcgctt gttacctttt tccctttttc 2400ccttttttcc ttattttggt tctttgaaaa ccacccgtgt gtgattctac gactgtgccc 2460gtttttacct atttccttac atttacgtgg acttctttct gcctt 2505531275DNAAspergillus nigerCDS(1)..(1275) 53atg ggt cag acc ttg tcc gag ccc gtg gtc gat aag act tcc gct gaa 48Met Gly Gln Thr Leu Ser Glu Pro Val Val Asp Lys Thr Ser Ala Glu 1 5 10 15 ggt caa gat gag tgc tgt ata tac ggt gtt tcc gcc atg caa gga tgg 96Gly Gln Asp Glu Cys Cys Ile Tyr Gly Val Ser Ala Met Gln Gly Trp 20 25 30 cga atc agc atg gag gat gcc cat gcc gct gtc ctc gac ctc cag gcc 144Arg Ile Ser Met Glu Asp Ala His Ala Ala Val Leu Asp Leu Gln Ala 35 40 45 aag tac tcc gag cag gat gaa aag ccg acc gac ccc gat aaa cga ctc 192Lys Tyr Ser Glu Gln Asp Glu Lys Pro Thr Asp Pro Asp Lys Arg Leu 50 55 60 gct ttc ttt ggt gta tat gac ggg cac ggt gga gac aaa gta gca tta 240Ala Phe Phe Gly Val Tyr Asp Gly His Gly Gly Asp Lys Val Ala Leu 65 70 75 80 ttc gcc gga gaa aac gtc cac aag atc gtc gcg aag cag gac tcc ttt 288Phe Ala Gly Glu Asn Val His Lys Ile Val Ala Lys Gln Asp Ser Phe 85 90 95 gcc aaa ggt gat atc gaa cag gcc ctg aag gat ggc ttc ctc gct acc 336Ala Lys Gly Asp Ile Glu Gln Ala Leu Lys Asp Gly Phe Leu Ala Thr 100 105 110 gac cgg gct att ttg gaa gac ccg aaa tat gag gag gaa gtg tct ggc 384Asp Arg Ala Ile Leu Glu Asp Pro Lys Tyr Glu Glu Glu Val Ser Gly 115 120 125 tgc acc gca gcc gtc agc gtt atc tcg aag cac aag atc tgg gtg gcc 432Cys Thr Ala Ala Val Ser Val Ile Ser Lys His Lys Ile Trp Val Ala 130 135 140 aat gct ggt gat tct cgc tct gta ctg ggt gtc aag ggt cgc gca aag 480Asn Ala Gly Asp Ser Arg Ser Val Leu Gly Val Lys Gly Arg Ala Lys 145 150 155 160 cct ctg tca ttt gac cac aag cct cag aat gaa ggc gag aaa gcc cgt 528Pro Leu Ser Phe Asp His Lys Pro Gln Asn Glu Gly Glu Lys Ala Arg 165 170 175 atc agc gct gct ggt ggt ttc gtt gac ttc ggc cgt gtc aac ggc aac 576Ile Ser Ala Ala Gly Gly Phe Val Asp Phe Gly Arg Val Asn Gly Asn 180 185 190 ctg gcc ttg tcg cgg gcc att ggt gac ttc gag ttc aag aag agc ccc 624Leu Ala Leu Ser Arg Ala Ile Gly Asp Phe Glu Phe Lys Lys Ser Pro 195 200 205 gag ttg tct cct gag cag cag atc gtc act gcc tat ccc gac gtc act 672Glu Leu Ser Pro Glu Gln Gln Ile Val Thr Ala Tyr Pro Asp Val Thr 210 215 220 gtg cac gat ctc agt gac gat gac gag ttc ctc gta att gcc tgt gac 720Val His Asp Leu Ser Asp Asp Asp Glu Phe Leu Val Ile Ala Cys Asp 225 230 235 240 ggt atc tgg gat tgt cag tcc tcc caa tca gtg gtg gaa ttc gtc cgc 768Gly Ile Trp Asp Cys Gln Ser Ser Gln Ser Val Val Glu Phe Val Arg 245 250 255 cgt ggg att gcc gcg aag cag gat ctg tat cgg atc tgt gag aac atg 816Arg Gly Ile Ala Ala Lys Gln Asp Leu Tyr Arg Ile Cys Glu Asn Met 260 265 270 atg gac aac tgc ctg gcc tcc aac agt gaa acc ggc ggc gtt ggc tgc 864Met Asp Asn Cys Leu Ala Ser Asn Ser Glu Thr Gly Gly Val Gly Cys 275 280 285 gac aat atg aca atg gtc atc atc ggt ctc ctg aac gct gag ttc cgc 912Asp Asn Met Thr Met Val Ile Ile Gly Leu Leu Asn Ala Glu Phe Arg 290 295 300 ggc ccc ggt atc cgg aat caa ttc gag gag aac ccg gat gac ttt gac 960Gly Pro Gly Ile Arg Asn Gln Phe Glu Glu Asn Pro Asp Asp Phe Asp 305 310 315 320 atg gaa aac gac cgt gcg cgt ggc ttc agc gtc cgc tcg ggc cgc atc 1008Met Glu Asn Asp Arg Ala Arg Gly Phe Ser Val Arg Ser Gly Arg Ile 325 330 335 atc ctc ttg ggg gac ggc act gaa ttg att ccg gag cag aac gat gac 1056Ile Leu Leu Gly Asp Gly Thr Glu Leu Ile Pro Glu Gln Asn Asp Asp 340 345 350 gaa ctc ttt gat cag gct gag gaa gac cag gac ctt gtc aat cag gtg 1104Glu Leu Phe Asp Gln Ala Glu Glu Asp Gln Asp Leu Val Asn Gln Val 355 360 365 cac cgt gat tcg cct gat gcg gct cgg aat gaa cgg gag gga act cct 1152His Arg Asp Ser Pro Asp Ala Ala Arg Asn Glu Arg Glu Gly Thr Pro 370 375 380 ggg cct cag tct aag gat act tct cga acg gac gcc gct gag ata tcg 1200Gly Pro Gln Ser Lys Asp Thr Ser Arg Thr Asp Ala Ala Glu Ile Ser 385 390 395 400 gag tcg ccg tct acc acc gcg gag ggt tcg tcc ggc agt ggc cct gga 1248Glu Ser Pro Ser Thr Thr Ala Glu Gly Ser Ser Gly Ser Gly Pro Gly 405 410 415 acg ccg caa aag cct acg agt tcg tag 1275Thr Pro Gln Lys Pro Thr Ser Ser 420 54424PRTAspergillus niger 54Met Gly Gln Thr Leu Ser Glu Pro Val Val Asp Lys Thr Ser Ala Glu 1 5 10 15 Gly Gln Asp Glu Cys Cys Ile Tyr Gly Val Ser Ala Met Gln Gly Trp 20 25 30 Arg Ile Ser Met Glu Asp Ala His Ala Ala Val Leu Asp Leu Gln Ala 35 40 45 Lys Tyr Ser Glu Gln Asp Glu Lys Pro Thr Asp Pro Asp Lys Arg Leu 50

55 60 Ala Phe Phe Gly Val Tyr Asp Gly His Gly Gly Asp Lys Val Ala Leu 65 70 75 80 Phe Ala Gly Glu Asn Val His Lys Ile Val Ala Lys Gln Asp Ser Phe 85 90 95 Ala Lys Gly Asp Ile Glu Gln Ala Leu Lys Asp Gly Phe Leu Ala Thr 100 105 110 Asp Arg Ala Ile Leu Glu Asp Pro Lys Tyr Glu Glu Glu Val Ser Gly 115 120 125 Cys Thr Ala Ala Val Ser Val Ile Ser Lys His Lys Ile Trp Val Ala 130 135 140 Asn Ala Gly Asp Ser Arg Ser Val Leu Gly Val Lys Gly Arg Ala Lys 145 150 155 160 Pro Leu Ser Phe Asp His Lys Pro Gln Asn Glu Gly Glu Lys Ala Arg 165 170 175 Ile Ser Ala Ala Gly Gly Phe Val Asp Phe Gly Arg Val Asn Gly Asn 180 185 190 Leu Ala Leu Ser Arg Ala Ile Gly Asp Phe Glu Phe Lys Lys Ser Pro 195 200 205 Glu Leu Ser Pro Glu Gln Gln Ile Val Thr Ala Tyr Pro Asp Val Thr 210 215 220 Val His Asp Leu Ser Asp Asp Asp Glu Phe Leu Val Ile Ala Cys Asp 225 230 235 240 Gly Ile Trp Asp Cys Gln Ser Ser Gln Ser Val Val Glu Phe Val Arg 245 250 255 Arg Gly Ile Ala Ala Lys Gln Asp Leu Tyr Arg Ile Cys Glu Asn Met 260 265 270 Met Asp Asn Cys Leu Ala Ser Asn Ser Glu Thr Gly Gly Val Gly Cys 275 280 285 Asp Asn Met Thr Met Val Ile Ile Gly Leu Leu Asn Ala Glu Phe Arg 290 295 300 Gly Pro Gly Ile Arg Asn Gln Phe Glu Glu Asn Pro Asp Asp Phe Asp 305 310 315 320 Met Glu Asn Asp Arg Ala Arg Gly Phe Ser Val Arg Ser Gly Arg Ile 325 330 335 Ile Leu Leu Gly Asp Gly Thr Glu Leu Ile Pro Glu Gln Asn Asp Asp 340 345 350 Glu Leu Phe Asp Gln Ala Glu Glu Asp Gln Asp Leu Val Asn Gln Val 355 360 365 His Arg Asp Ser Pro Asp Ala Ala Arg Asn Glu Arg Glu Gly Thr Pro 370 375 380 Gly Pro Gln Ser Lys Asp Thr Ser Arg Thr Asp Ala Ala Glu Ile Ser 385 390 395 400 Glu Ser Pro Ser Thr Thr Ala Glu Gly Ser Ser Gly Ser Gly Pro Gly 405 410 415 Thr Pro Gln Lys Pro Thr Ser Ser 420 551209DNAAspergillus niger 55ctggcagtta tttagtggtg attcggcatc atccccttat cgatcatact cgcccgtctt 60ctctcgagtc cttaaacgcc aaaagacgac tgtctgcatc ctctctattt cgcttaccgc 120ttcgtcgcat cgtacccgcc acccgagcaa cctcccccct aagttaatcc caacgttcgc 180aactctacta cccatcaatt atggccgcca tctggggtaa cggcgggcag gctggccagt 240tcccgctgga gcaatggttc tatgaaatgc cccctgtaac tcgatggtgg acagcagcca 300cagttgccac ttcagtcttg gtccaatgtc acgtcctcac cccattccag ctgttttata 360gcttccgcgc agtctatgtt aagtctcagg tacgtcgcag ctagtacttc cgtccactgt 420atagggtaga cgaatcacgc ggctaaccat cgcatagtat tggcgtctgt tcacaacctt 480cctatacttc ggaccactca atctcgactt actatttcat gtgttcttct tgcagcgata 540ctcgcgcctc ttggaggaat catcggggcg atcgccggcc cacttctcgt ggcttctgtt 600ctacgccatg gcctctctcc tcgtcctctc gccatttctc tcccttccat tcctgggcac 660ggctctctct tccagtctgg tctacatctg gagtcgtcgc aacccggaaa ctcgcctcag 720cttcctagga atgctggtct tcaccgcccc ctatctcccc tgggttctga tggcattcag 780cctggtcgtc catggcatcg tgcccaagga tgaaatctgc ggcgttgtcg tcggccacgt 840ctggtacttc ttcaacgatg tttacccttc gcttcacggt ggtcaccgtc ctttcgatcc 900tcctatgtgg tgggtgcgtc tgtttgagtc agggcccggg gaacgaggca ccgacgctgc 960caacgtcaac ggggaattcg ccgctgctgc tgcacccgaa gttcggtgag ctatttgtgc 1020accccactgg ggcatttact gcatggcgat gcaaagaatc gtccgcgtaa tcgctctgga 1080aacgtcagca tatatgtgtg tactgccaac tactcgcgcc gacacgcgcg aagcatgaga 1140agttaatact gtcaggatat aagcaaggat cacggcggca gacttgatgg gatttcttat 1200cgtgtggct 120956741DNAAspergillus nigerCDS(1)..(741) 56atg gcc gcc atc tgg ggt aac ggc ggg cag gct ggc cag ttc ccg ctg 48Met Ala Ala Ile Trp Gly Asn Gly Gly Gln Ala Gly Gln Phe Pro Leu 1 5 10 15 gag caa tgg ttc tat gaa atg ccc cct gta act cga tgg tgg aca gca 96Glu Gln Trp Phe Tyr Glu Met Pro Pro Val Thr Arg Trp Trp Thr Ala 20 25 30 gcc aca gtt gcc act tca gtc ttg gtc caa tgt cac gtc ctc acc cca 144Ala Thr Val Ala Thr Ser Val Leu Val Gln Cys His Val Leu Thr Pro 35 40 45 ttc cag ctg ttt tat agc ttc cgc gca gtc tat gtt aag tct cag tat 192Phe Gln Leu Phe Tyr Ser Phe Arg Ala Val Tyr Val Lys Ser Gln Tyr 50 55 60 tgg cgt ctg ttc aca acc ttc cta tac ttc gga cca ctc aat ctc gac 240Trp Arg Leu Phe Thr Thr Phe Leu Tyr Phe Gly Pro Leu Asn Leu Asp 65 70 75 80 tta cta ttt cat gtg ttc ttc ttg cag cga tac tcg cgc ctc ttg gag 288Leu Leu Phe His Val Phe Phe Leu Gln Arg Tyr Ser Arg Leu Leu Glu 85 90 95 gaa tca tcg ggg cga tcg ccg gcc cac ttc tcg tgg ctt ctg ttc tac 336Glu Ser Ser Gly Arg Ser Pro Ala His Phe Ser Trp Leu Leu Phe Tyr 100 105 110 gcc atg gcc tct ctc ctc gtc ctc tcg cca ttt ctc tcc ctt cca ttc 384Ala Met Ala Ser Leu Leu Val Leu Ser Pro Phe Leu Ser Leu Pro Phe 115 120 125 ctg ggc acg gct ctc tct tcc agt ctg gtc tac atc tgg agt cgt cgc 432Leu Gly Thr Ala Leu Ser Ser Ser Leu Val Tyr Ile Trp Ser Arg Arg 130 135 140 aac ccg gaa act cgc ctc agc ttc cta gga atg ctg gtc ttc acc gcc 480Asn Pro Glu Thr Arg Leu Ser Phe Leu Gly Met Leu Val Phe Thr Ala 145 150 155 160 ccc tat ctc ccc tgg gtt ctg atg gca ttc agc ctg gtc gtc cat ggc 528Pro Tyr Leu Pro Trp Val Leu Met Ala Phe Ser Leu Val Val His Gly 165 170 175 atc gtg ccc aag gat gaa atc tgc ggc gtt gtc gtc ggc cac gtc tgg 576Ile Val Pro Lys Asp Glu Ile Cys Gly Val Val Val Gly His Val Trp 180 185 190 tac ttc ttc aac gat gtt tac cct tcg ctt cac ggt ggt cac cgt cct 624Tyr Phe Phe Asn Asp Val Tyr Pro Ser Leu His Gly Gly His Arg Pro 195 200 205 ttc gat cct cct atg tgg tgg gtg cgt ctg ttt gag tca ggg ccc ggg 672Phe Asp Pro Pro Met Trp Trp Val Arg Leu Phe Glu Ser Gly Pro Gly 210 215 220 gaa cga ggc acc gac gct gcc aac gtc aac ggg gaa ttc gcc gct gct 720Glu Arg Gly Thr Asp Ala Ala Asn Val Asn Gly Glu Phe Ala Ala Ala 225 230 235 240 gct gca ccc gaa gtt cgg tga 741Ala Ala Pro Glu Val Arg 245 57246PRTAspergillus niger 57Met Ala Ala Ile Trp Gly Asn Gly Gly Gln Ala Gly Gln Phe Pro Leu 1 5 10 15 Glu Gln Trp Phe Tyr Glu Met Pro Pro Val Thr Arg Trp Trp Thr Ala 20 25 30 Ala Thr Val Ala Thr Ser Val Leu Val Gln Cys His Val Leu Thr Pro 35 40 45 Phe Gln Leu Phe Tyr Ser Phe Arg Ala Val Tyr Val Lys Ser Gln Tyr 50 55 60 Trp Arg Leu Phe Thr Thr Phe Leu Tyr Phe Gly Pro Leu Asn Leu Asp 65 70 75 80 Leu Leu Phe His Val Phe Phe Leu Gln Arg Tyr Ser Arg Leu Leu Glu 85 90 95 Glu Ser Ser Gly Arg Ser Pro Ala His Phe Ser Trp Leu Leu Phe Tyr 100 105 110 Ala Met Ala Ser Leu Leu Val Leu Ser Pro Phe Leu Ser Leu Pro Phe 115 120 125 Leu Gly Thr Ala Leu Ser Ser Ser Leu Val Tyr Ile Trp Ser Arg Arg 130 135 140 Asn Pro Glu Thr Arg Leu Ser Phe Leu Gly Met Leu Val Phe Thr Ala 145 150 155 160 Pro Tyr Leu Pro Trp Val Leu Met Ala Phe Ser Leu Val Val His Gly 165 170 175 Ile Val Pro Lys Asp Glu Ile Cys Gly Val Val Val Gly His Val Trp 180 185 190 Tyr Phe Phe Asn Asp Val Tyr Pro Ser Leu His Gly Gly His Arg Pro 195 200 205 Phe Asp Pro Pro Met Trp Trp Val Arg Leu Phe Glu Ser Gly Pro Gly 210 215 220 Glu Arg Gly Thr Asp Ala Ala Asn Val Asn Gly Glu Phe Ala Ala Ala 225 230 235 240 Ala Ala Pro Glu Val Arg 245 582538DNAAspergillus niger 58gaagcttggc ggtggtggtc atgtctttcc aggcccctcg gaacagttct ccattttcgt 60ctcagcaacc gttccagaat aactactggc gtgctagtcg aagccctggg accaacgggc 120tgcctggcta tgggttttcg ccaccgactg gtatctccaa ctccctgaac aatcccctcg 180ccggcgaccg cactctcccc atgtacaagg acaaacctta cttcgcacca cgacgcaccg 240gtccgagagc caggcggcgg aagatcatat atagtgggct atgtctgttc gtcctgctcg 300ctctgtggta ctactctggc tctggtaagc cggaatggaa gacaccggac gcggagaagg 360gcgccgagct ctggaagtgg gtgcaaagtt ttgaggagtc ggaaccacca tacgatggca 420gcgcagcgac agagaagatt gactgggaag caaggaggga gaaagtgcgc gacgtcttca 480ttgtcagttg ggatggttat gcggctaatg cgtggggtga gtttctggct ggaattaacc 540tcttgattct tggatgtggt gactgacaat ggctttgtct gccggtgatt aggttacgat 600gaataccacc caattgccaa aaacggtcgg cacatgattg aaggaggaat ggggtggata 660attgttgatg cgctggacac tttgatgatt atgaacctga cgtcgcgagt gcaacatgcc 720cgcagctgga tccacaactc gttgcaatac aaccaggacc acgatgtgaa taccttcgag 780actaccattc gcatgctggg aggcttactc tccgcacact atctctccac gaactatccc 840gagctagctc cgcttacgga tgacgataca ggcgcgccgg gagaagactt gtatatcgag 900aaggccaccg atctggcaga tcgtctattg ggtgcttttg aatccggcac tggaatcccg 960tatgcaagca tcaatttgaa caaatccgag ggccttccct cgtacgcgga taatggcgcc 1020tcatctactg ccgaagccac tactctccag ctggagttca agtacttggc caaactgacg 1080ggcgaggccg agtactggca ggctgtggag aaggttatgg aggtcgtgga cgaccagaag 1140atggaagacg gattgcttcc gatctacgta tatccagaga ccggcgaatt taaaggcgat 1200aacatccgtc tcggcagtag aggcgattca tactatggta tacatgatgg ctttgtgcgt 1260gaatattcaa tgcgctgacc gatgtttcct agaatacctc atcaagcagt accttcaaac 1320gcaagagact gaaccgatct acaaggacat gtgggatgag tccctcgtcg gcgtgcgcaa 1380gcacctgatc acctatacac agaacgccaa gttgaccgtt ctgggcgaac gccctgccgg 1440gctgcatgga gtcctttccc ccaagatgga ccacctagtc tgcttctacc ccggcaccat 1500cgctctcgca gcaactggcg gacgtcctct gtccgaggca agacagtcac ccgattgggg 1560ccaacgccag gaggaagaga ttcttctagc ccgagaacta accaagacct gctgggcaac 1620gtatctaatc acgaagaccg gcctcgcacc agagatcacc tacttcaatg tcgacgaccc 1680tcgcgtcatg gaaacagaca tgtacccaga ctcgaccatt gccaaaccca gctcaggcca 1740gcaaaaagcc tctggcgaac tccccctcct ctccaaatcc atctaccccg tcagcgacta 1800ctccaccaaa tggcgcgacg acctcaacat ccacaaacaa gaccgccaca acctccaacg 1860cccagagacc gtcgaatccc tcttctacat gtatcgcatc accggagacg acatctaccg 1920acactggggc tgggagatgt tcaagtcttt tgtcaagcat actgccgtgg tcgaggacat 1980ccctgtcgat gaactcagca aagaagacgc gtccagctca acatcatcat cggaggaaga 2040agacggcgga actcagaagc caaaacctca gaaaatcacg ggcttcacct cgctctcaaa 2100cgctgatgac aaccctccag tgaagcgcga caacatggag agtttctgga tggccgagac 2160gcttaaatac ttctacttgc tattctcgga ccgcgacttt atctctctcg aagaccatgt 2220attcaatacg gaggcgcatc ctctcccgcg gttcaagcca acgggcgagt tgaagaccgg 2280gtggatgagg aagagtcgga cgataccaac atccagtgag gtggaggagt cggtataatt 2340tctttgacta tctatttttt ttcttatgct tttatatatc tctctcttgg actacgtctc 2400cttagatccc tctaacaaaa aagaaagata tctcgaattg attgatcatc tcttgatttt 2460gtttgtttga tgcttgctta tatgtactgc tatatattgc tcctgcattc tggctggtta 2520catccaaggc agggactt 2538591812DNAAspergillus nigerCDS(1)..(1812) 59atg tac aag gac aaa cct tac ttc gca cca cga cgc acc ggt ccg aga 48Met Tyr Lys Asp Lys Pro Tyr Phe Ala Pro Arg Arg Thr Gly Pro Arg 1 5 10 15 gcc agg cgg cgg aag atc ata tat agt ggg cta tgt ctg ttc gtc ctg 96Ala Arg Arg Arg Lys Ile Ile Tyr Ser Gly Leu Cys Leu Phe Val Leu 20 25 30 ctc gct ctg tgg tac tac tct ggc tct ggt aag ccg gaa tgg aag aca 144Leu Ala Leu Trp Tyr Tyr Ser Gly Ser Gly Lys Pro Glu Trp Lys Thr 35 40 45 ccg gac gcg gag aag ggc gcc gag ctc tgg aag tgg gtg caa agt ttt 192Pro Asp Ala Glu Lys Gly Ala Glu Leu Trp Lys Trp Val Gln Ser Phe 50 55 60 gag gag tcg gaa cca cca tac gat ggc agc gca gcg aca gag aag att 240Glu Glu Ser Glu Pro Pro Tyr Asp Gly Ser Ala Ala Thr Glu Lys Ile 65 70 75 80 gac tgg gaa gca agg agg gag aaa gtg cgc gac gtc ttc att gtc agt 288Asp Trp Glu Ala Arg Arg Glu Lys Val Arg Asp Val Phe Ile Val Ser 85 90 95 tgg gat ggt tat gcg gct aat gcg tgg ggt tac gat gaa tac cac cca 336Trp Asp Gly Tyr Ala Ala Asn Ala Trp Gly Tyr Asp Glu Tyr His Pro 100 105 110 att gcc aaa aac ggt cgg cac atg att gaa gga gga atg ggg tgg ata 384Ile Ala Lys Asn Gly Arg His Met Ile Glu Gly Gly Met Gly Trp Ile 115 120 125 att gtt gat gcg ctg gac act ttg atg att atg aac ctg acg tcg cga 432Ile Val Asp Ala Leu Asp Thr Leu Met Ile Met Asn Leu Thr Ser Arg 130 135 140 gtg caa cat gcc cgc agc tgg atc cac aac tcg ttg caa tac aac cag 480Val Gln His Ala Arg Ser Trp Ile His Asn Ser Leu Gln Tyr Asn Gln 145 150 155 160 gac cac gat gtg aat acc ttc gag act acc att cgc atg ctg gga ggc 528Asp His Asp Val Asn Thr Phe Glu Thr Thr Ile Arg Met Leu Gly Gly 165 170 175 tta ctc tcc gca cac tat ctc tcc acg aac tat ccc gag cta gct ccg 576Leu Leu Ser Ala His Tyr Leu Ser Thr Asn Tyr Pro Glu Leu Ala Pro 180 185 190 ctt acg gat gac gat aca ggc gcg ccg gga gaa gac ttg tat atc gag 624Leu Thr Asp Asp Asp Thr Gly Ala Pro Gly Glu Asp Leu Tyr Ile Glu 195 200 205 aag gcc acc gat ctg gca gat cgt cta ttg ggt gct ttt gaa tcc ggc 672Lys Ala Thr Asp Leu Ala Asp Arg Leu Leu Gly Ala Phe Glu Ser Gly 210 215 220 act gga atc ccg tat gca agc atc aat ttg aac aaa tcc gag ggc ctt 720Thr Gly Ile Pro Tyr Ala Ser Ile Asn Leu Asn Lys Ser Glu Gly Leu 225 230 235 240 ccc tcg tac gcg gat aat ggc gcc tca tct act gcc gaa gcc act act 768Pro Ser Tyr Ala Asp Asn Gly Ala Ser Ser Thr Ala Glu Ala Thr Thr 245 250 255 ctc cag ctg gag ttc aag tac ttg gcc aaa ctg acg ggc gag gcc gag 816Leu Gln Leu Glu Phe Lys Tyr Leu Ala Lys Leu Thr Gly Glu Ala Glu 260 265 270 tac tgg cag gct gtg gag aag gtt atg gag gtc gtg gac gac cag aag 864Tyr Trp Gln Ala Val Glu Lys Val Met Glu Val Val Asp Asp Gln Lys 275 280 285 atg gaa gac gga ttg ctt ccg atc tac gta tat cca gag acc ggc gaa 912Met Glu Asp Gly Leu Leu Pro Ile Tyr Val Tyr Pro Glu Thr Gly Glu 290 295 300 ttt aaa ggc gat aac atc cgt ctc ggc agt aga ggc gat tca tac tat 960Phe Lys Gly Asp Asn Ile Arg Leu Gly Ser Arg Gly Asp Ser Tyr Tyr 305 310 315 320 gaa tac ctc atc aag cag tac ctt caa acg caa gag act gaa ccg atc 1008Glu Tyr Leu Ile Lys Gln Tyr Leu Gln Thr Gln Glu Thr Glu Pro Ile 325 330 335 tac aag gac atg tgg gat gag tcc ctc gtc ggc gtg cgc aag cac ctg 1056Tyr Lys Asp Met Trp Asp Glu Ser Leu Val Gly Val Arg Lys His Leu 340 345 350 atc acc tat aca cag aac gcc aag ttg acc gtt ctg ggc gaa cgc cct 1104Ile Thr Tyr Thr Gln Asn Ala Lys Leu Thr Val Leu Gly Glu Arg Pro 355 360 365 gcc ggg ctg cat gga gtc ctt tcc ccc aag atg gac cac cta gtc tgc 1152Ala Gly Leu His Gly Val Leu Ser Pro Lys Met Asp His Leu Val Cys 370 375 380 ttc tac ccc ggc acc atc gct ctc gca gca act ggc gga cgt cct ctg 1200Phe Tyr Pro Gly Thr Ile Ala Leu Ala Ala Thr Gly Gly Arg Pro Leu 385 390 395 400 tcc gag gca aga cag tca ccc gat tgg ggc caa cgc cag gag gaa gag 1248Ser Glu Ala Arg Gln Ser Pro Asp Trp Gly Gln Arg Gln Glu Glu Glu 405 410 415 att ctt cta gcc cga gaa cta acc aag acc tgc tgg gca acg tat cta 1296Ile Leu Leu Ala Arg Glu Leu Thr Lys Thr Cys Trp Ala Thr Tyr Leu 420 425 430 atc acg aag acc ggc ctc gca cca gag

atc acc tac ttc aat gtc gac 1344Ile Thr Lys Thr Gly Leu Ala Pro Glu Ile Thr Tyr Phe Asn Val Asp 435 440 445 gac cct cgc gtc atg gaa aca gac atc gac tac tcc acc aaa tgg cgc 1392Asp Pro Arg Val Met Glu Thr Asp Ile Asp Tyr Ser Thr Lys Trp Arg 450 455 460 gac gac ctc aac atc cac aaa caa gac cgc cac aac ctc caa cgc cca 1440Asp Asp Leu Asn Ile His Lys Gln Asp Arg His Asn Leu Gln Arg Pro 465 470 475 480 gag acc gtc gaa tcc ctc ttc tac atg tat cgc atc acc gga gac gac 1488Glu Thr Val Glu Ser Leu Phe Tyr Met Tyr Arg Ile Thr Gly Asp Asp 485 490 495 atc tac cga cac tgg ggc tgg gag atg ttc aag tct ttt gtc aag cat 1536Ile Tyr Arg His Trp Gly Trp Glu Met Phe Lys Ser Phe Val Lys His 500 505 510 act gcc gtg aaa atc acg ggc ttc acc tcg ctc tca aac gct gat gac 1584Thr Ala Val Lys Ile Thr Gly Phe Thr Ser Leu Ser Asn Ala Asp Asp 515 520 525 aac cct cca gtg aag cgc gac aac atg gag agt ttc tgg atg gcc gag 1632Asn Pro Pro Val Lys Arg Asp Asn Met Glu Ser Phe Trp Met Ala Glu 530 535 540 acg ctt aaa tac ttc tac ttg cta ttc tcg gac cgc gac ttt atc tct 1680Thr Leu Lys Tyr Phe Tyr Leu Leu Phe Ser Asp Arg Asp Phe Ile Ser 545 550 555 560 ctc gaa gac cat gta ttc aat acg gag gcg cat cct ctc ccg cgg ttc 1728Leu Glu Asp His Val Phe Asn Thr Glu Ala His Pro Leu Pro Arg Phe 565 570 575 aag cca acg ggc gag ttg aag acc ggg tgg atg agg aag agt cgg acg 1776Lys Pro Thr Gly Glu Leu Lys Thr Gly Trp Met Arg Lys Ser Arg Thr 580 585 590 ata cca aca tcc agt gag gtg gag gag tcg gta taa 1812Ile Pro Thr Ser Ser Glu Val Glu Glu Ser Val 595 600 60603PRTAspergillus niger 60Met Tyr Lys Asp Lys Pro Tyr Phe Ala Pro Arg Arg Thr Gly Pro Arg 1 5 10 15 Ala Arg Arg Arg Lys Ile Ile Tyr Ser Gly Leu Cys Leu Phe Val Leu 20 25 30 Leu Ala Leu Trp Tyr Tyr Ser Gly Ser Gly Lys Pro Glu Trp Lys Thr 35 40 45 Pro Asp Ala Glu Lys Gly Ala Glu Leu Trp Lys Trp Val Gln Ser Phe 50 55 60 Glu Glu Ser Glu Pro Pro Tyr Asp Gly Ser Ala Ala Thr Glu Lys Ile 65 70 75 80 Asp Trp Glu Ala Arg Arg Glu Lys Val Arg Asp Val Phe Ile Val Ser 85 90 95 Trp Asp Gly Tyr Ala Ala Asn Ala Trp Gly Tyr Asp Glu Tyr His Pro 100 105 110 Ile Ala Lys Asn Gly Arg His Met Ile Glu Gly Gly Met Gly Trp Ile 115 120 125 Ile Val Asp Ala Leu Asp Thr Leu Met Ile Met Asn Leu Thr Ser Arg 130 135 140 Val Gln His Ala Arg Ser Trp Ile His Asn Ser Leu Gln Tyr Asn Gln 145 150 155 160 Asp His Asp Val Asn Thr Phe Glu Thr Thr Ile Arg Met Leu Gly Gly 165 170 175 Leu Leu Ser Ala His Tyr Leu Ser Thr Asn Tyr Pro Glu Leu Ala Pro 180 185 190 Leu Thr Asp Asp Asp Thr Gly Ala Pro Gly Glu Asp Leu Tyr Ile Glu 195 200 205 Lys Ala Thr Asp Leu Ala Asp Arg Leu Leu Gly Ala Phe Glu Ser Gly 210 215 220 Thr Gly Ile Pro Tyr Ala Ser Ile Asn Leu Asn Lys Ser Glu Gly Leu 225 230 235 240 Pro Ser Tyr Ala Asp Asn Gly Ala Ser Ser Thr Ala Glu Ala Thr Thr 245 250 255 Leu Gln Leu Glu Phe Lys Tyr Leu Ala Lys Leu Thr Gly Glu Ala Glu 260 265 270 Tyr Trp Gln Ala Val Glu Lys Val Met Glu Val Val Asp Asp Gln Lys 275 280 285 Met Glu Asp Gly Leu Leu Pro Ile Tyr Val Tyr Pro Glu Thr Gly Glu 290 295 300 Phe Lys Gly Asp Asn Ile Arg Leu Gly Ser Arg Gly Asp Ser Tyr Tyr 305 310 315 320 Glu Tyr Leu Ile Lys Gln Tyr Leu Gln Thr Gln Glu Thr Glu Pro Ile 325 330 335 Tyr Lys Asp Met Trp Asp Glu Ser Leu Val Gly Val Arg Lys His Leu 340 345 350 Ile Thr Tyr Thr Gln Asn Ala Lys Leu Thr Val Leu Gly Glu Arg Pro 355 360 365 Ala Gly Leu His Gly Val Leu Ser Pro Lys Met Asp His Leu Val Cys 370 375 380 Phe Tyr Pro Gly Thr Ile Ala Leu Ala Ala Thr Gly Gly Arg Pro Leu 385 390 395 400 Ser Glu Ala Arg Gln Ser Pro Asp Trp Gly Gln Arg Gln Glu Glu Glu 405 410 415 Ile Leu Leu Ala Arg Glu Leu Thr Lys Thr Cys Trp Ala Thr Tyr Leu 420 425 430 Ile Thr Lys Thr Gly Leu Ala Pro Glu Ile Thr Tyr Phe Asn Val Asp 435 440 445 Asp Pro Arg Val Met Glu Thr Asp Ile Asp Tyr Ser Thr Lys Trp Arg 450 455 460 Asp Asp Leu Asn Ile His Lys Gln Asp Arg His Asn Leu Gln Arg Pro 465 470 475 480 Glu Thr Val Glu Ser Leu Phe Tyr Met Tyr Arg Ile Thr Gly Asp Asp 485 490 495 Ile Tyr Arg His Trp Gly Trp Glu Met Phe Lys Ser Phe Val Lys His 500 505 510 Thr Ala Val Lys Ile Thr Gly Phe Thr Ser Leu Ser Asn Ala Asp Asp 515 520 525 Asn Pro Pro Val Lys Arg Asp Asn Met Glu Ser Phe Trp Met Ala Glu 530 535 540 Thr Leu Lys Tyr Phe Tyr Leu Leu Phe Ser Asp Arg Asp Phe Ile Ser 545 550 555 560 Leu Glu Asp His Val Phe Asn Thr Glu Ala His Pro Leu Pro Arg Phe 565 570 575 Lys Pro Thr Gly Glu Leu Lys Thr Gly Trp Met Arg Lys Ser Arg Thr 580 585 590 Ile Pro Thr Ser Ser Glu Val Glu Glu Ser Val 595 600 612113DNAAspergillus niger 61cccgcaatcc ccgtcgacct catcgcttcc tccctttctc ctccatcctc tctctcttcc 60gtcgtctttt cttcttctcc ttctcctttt gtacttcccc tccattcctt cagctggttc 120tcgcctccag ctttcctttc tttctttccc tcccctttta ttcgagtaat cctgcagctc 180tgggaggtgc aacagtcaca atgagcggac gtgagtcttg cacgcgatcg ctgccatctc 240cgcgacagcg ttccatcctt tacctcaatg gatcagcaaa tgctgatact cgattctagt 300ccggtttctc gatctcatca agcccttcac gcccctcctc ccggaggtgg ccgccccgga 360aaccaaggtt cccttcaacc agaagttgat gtggacgggg gtacgtgata cttgtccagc 420tcgacatgag cttctaagct aatggattac ccctgcagtt gaccctattg atcttcctgg 480tcatgagcca gatgcccttg tacggaattg tctcctctga cacctccgac cctctgtact 540ggctccgtat gatgttggcc agtaaccggg gtaccctgat ggaactgggt atcaccccca 600tcatctcctc tggcatggtt ttccaggtat gtaatgggga aattgcaatc tgatcacgga 660tatcgggcat ttgctaatat gtggcttttg tctgatagct tctcgctggt acccacctca 720tcgatgtcaa cctggacctg aagaccgacc gtgaactgta tcagaccgct cagaagctct 780tcgctatcat cctgtccttc ggtcaggcct gcgtctacgt cctcactggt ctttacggcc 840agcccagtga ccttggtgcc ggtatctgtg ttctgctgat tgttcagctg gtcgttgctg 900gcttggttgt catcctgctg gatgagctgc tccagaaggg ctatggtctt ggtagcggta 960tctctctgtt catcgcgacc aacatctgcg agtcgatcgt ctggaaggct ttctctccta 1020cgaccatcaa cactggccgt ggtcccgagt ttgagggtgc catcattgcc ctcttccacc 1080ttctgttgac ctggtccgac aagcagcgcg ctctccgcga ggctttctac cgccagaacc 1140tccccaacat catgaacctg ctggctactc tcctcgtttt cgccgctgtg atctacctcc 1200agggcttccg tgttgagatc cctgtcaagt cctcccgcca gcgtggcatg cgtggttcct 1260accctgttcg cctgttctac acctccaaca tgcccatcat gcttcagtct gctctgtgct 1320ccaacatctt cctcatcagt cagatgctgt actctcgctt ctctgacaac ctccttgtca 1380agcttctcgg tgtttgggag cctcgtgagg gttctgccca gctccacgcc gcctccggca 1440ttgcctacta catgtctcct cccctgaact tcaaggaggc ccttcttgac cccattcaca 1500ccgccgttta catcaccttc atgctggttg cttgtgctct cttctggaag acctggattg 1560aggtttccgg ctctgctccc cgcgatgttg ccaagcagct caaggaccag ggtctcgtga 1620tggctggtca ccgtgagcag agcatgtaca aggagctcaa gcgcgtcatc cctactgctg 1680ctgctttcgg tggtgcctgc attggtgccc tgtccgtcgc ttctgacctg cttggtgctc 1740ttggcagcgg tactggtatc ctccttgccg ttacgtaagt cttcactttg gtctcagatt 1800ttctgaagtg gatactaaca ttcaaatgca ggattatata cggatacttt gaaattgccg 1860cccgtgaggg cgacattgga tcgggcctca agggccttgt tccgggtaac tagataaggc 1920cccctttttg atgaaagcat gagaagaagt ttgagggctt atgtttgttc ttgcaacttt 1980ctgtttcttc tcaggtagtg tgctgttgtg gctgggatct ggattattta gtttcttgat 2040ggatgtatgg ctagttttaa caatttgcag gaggggaaga tcttctctac ggagatacgt 2100ccacgccaca gct 2113621437DNAAspergillus nigerCDS(1)..(1437) 62atg agc gga ctc cgg ttt ctc gat ctc atc aag ccc ttc acg ccc ctc 48Met Ser Gly Leu Arg Phe Leu Asp Leu Ile Lys Pro Phe Thr Pro Leu 1 5 10 15 ctc ccg gag gtg gcc gcc ccg gaa acc aag gtt ccc ttc aac cag aag 96Leu Pro Glu Val Ala Ala Pro Glu Thr Lys Val Pro Phe Asn Gln Lys 20 25 30 ttg atg tgg acg ggg ttg acc cta ttg atc ttc ctg gtc atg agc cag 144Leu Met Trp Thr Gly Leu Thr Leu Leu Ile Phe Leu Val Met Ser Gln 35 40 45 atg ccc ttg tac gga att gtc tcc tct gac acc tcc gac cct ctg tac 192Met Pro Leu Tyr Gly Ile Val Ser Ser Asp Thr Ser Asp Pro Leu Tyr 50 55 60 tgg ctc cgt atg atg ttg gcc agt aac cgg ggt acc ctg atg gaa ctg 240Trp Leu Arg Met Met Leu Ala Ser Asn Arg Gly Thr Leu Met Glu Leu 65 70 75 80 ggt atc acc ccc atc atc tcc tct ggc atg gtt ttc cag ctt ctc gct 288Gly Ile Thr Pro Ile Ile Ser Ser Gly Met Val Phe Gln Leu Leu Ala 85 90 95 ggt acc cac ctc atc gat gtc aac ctg gac ctg aag acc gac cgt gaa 336Gly Thr His Leu Ile Asp Val Asn Leu Asp Leu Lys Thr Asp Arg Glu 100 105 110 ctg tat cag acc gct cag aag ctc ttc gct atc atc ctg tcc ttc ggt 384Leu Tyr Gln Thr Ala Gln Lys Leu Phe Ala Ile Ile Leu Ser Phe Gly 115 120 125 cag gcc tgc gtc tac gtc ctc act ggt ctt tac ggc cag ccc agt gac 432Gln Ala Cys Val Tyr Val Leu Thr Gly Leu Tyr Gly Gln Pro Ser Asp 130 135 140 ctt ggt gcc ggt atc tgt gtt ctg ctg att gtt cag ctg gtc gtt gct 480Leu Gly Ala Gly Ile Cys Val Leu Leu Ile Val Gln Leu Val Val Ala 145 150 155 160 ggc ttg gtt gtc atc ctg ctg gat gag ctg ctc cag aag ggc tat ggt 528Gly Leu Val Val Ile Leu Leu Asp Glu Leu Leu Gln Lys Gly Tyr Gly 165 170 175 ctt ggt agc ggt atc tct ctg ttc atc gcg acc aac atc tgc gag tcg 576Leu Gly Ser Gly Ile Ser Leu Phe Ile Ala Thr Asn Ile Cys Glu Ser 180 185 190 atc gtc tgg aag gct ttc tct cct acg acc atc aac act ggc cgt ggt 624Ile Val Trp Lys Ala Phe Ser Pro Thr Thr Ile Asn Thr Gly Arg Gly 195 200 205 ccc gag ttt gag ggt gcc atc att gcc ctc ttc cac ctt ctg ttg acc 672Pro Glu Phe Glu Gly Ala Ile Ile Ala Leu Phe His Leu Leu Leu Thr 210 215 220 tgg tcc gac aag cag cgc gct ctc cgc gag gct ttc tac cgc cag aac 720Trp Ser Asp Lys Gln Arg Ala Leu Arg Glu Ala Phe Tyr Arg Gln Asn 225 230 235 240 ctc ccc aac atc atg aac ctg ctg gct act ctc ctc gtt ttc gcc gct 768Leu Pro Asn Ile Met Asn Leu Leu Ala Thr Leu Leu Val Phe Ala Ala 245 250 255 gtg atc tac ctc cag ggc ttc cgt gtt gag atc cct gtc aag tcc tcc 816Val Ile Tyr Leu Gln Gly Phe Arg Val Glu Ile Pro Val Lys Ser Ser 260 265 270 cgc cag cgt ggc atg cgt ggt tcc tac cct gtt cgc ctg ttc tac acc 864Arg Gln Arg Gly Met Arg Gly Ser Tyr Pro Val Arg Leu Phe Tyr Thr 275 280 285 tcc aac atg ccc atc atg ctt cag tct gct ctg tgc tcc aac atc ttc 912Ser Asn Met Pro Ile Met Leu Gln Ser Ala Leu Cys Ser Asn Ile Phe 290 295 300 ctc atc agt cag atg ctg tac tct cgc ttc tct gac aac ctc ctt gtc 960Leu Ile Ser Gln Met Leu Tyr Ser Arg Phe Ser Asp Asn Leu Leu Val 305 310 315 320 aag ctt ctc ggt gtt tgg gag cct cgt gag ggt tct gcc cag ctc cac 1008Lys Leu Leu Gly Val Trp Glu Pro Arg Glu Gly Ser Ala Gln Leu His 325 330 335 gcc gcc tcc ggc att gcc tac tac atg tct cct ccc ctg aac ttc aag 1056Ala Ala Ser Gly Ile Ala Tyr Tyr Met Ser Pro Pro Leu Asn Phe Lys 340 345 350 gag gcc ctt ctt gac ccc att cac acc gcc gtt tac atc acc ttc atg 1104Glu Ala Leu Leu Asp Pro Ile His Thr Ala Val Tyr Ile Thr Phe Met 355 360 365 ctg gtt gct tgt gct ctc ttc tgg aag acc tgg att gag gtt tcc ggc 1152Leu Val Ala Cys Ala Leu Phe Trp Lys Thr Trp Ile Glu Val Ser Gly 370 375 380 tct gct ccc cgc gat gtt gcc aag cag ctc aag gac cag ggt ctc gtg 1200Ser Ala Pro Arg Asp Val Ala Lys Gln Leu Lys Asp Gln Gly Leu Val 385 390 395 400 atg gct ggt cac cgt gag cag agc atg tac aag gag ctc aag cgc gtc 1248Met Ala Gly His Arg Glu Gln Ser Met Tyr Lys Glu Leu Lys Arg Val 405 410 415 atc cct act gct gct gct ttc ggt ggt gcc tgc att ggt gcc ctg tcc 1296Ile Pro Thr Ala Ala Ala Phe Gly Gly Ala Cys Ile Gly Ala Leu Ser 420 425 430 gtc gct tct gac ctg ctt ggt gct ctt ggc agc ggt act ggt atc ctc 1344Val Ala Ser Asp Leu Leu Gly Ala Leu Gly Ser Gly Thr Gly Ile Leu 435 440 445 ctt gcc gtt acg att ata tac gga tac ttt gaa att gcc gcc cgt gag 1392Leu Ala Val Thr Ile Ile Tyr Gly Tyr Phe Glu Ile Ala Ala Arg Glu 450 455 460 ggc gac att gga tcg ggc ctc aag ggc ctt gtt ccg ggt aac tag 1437Gly Asp Ile Gly Ser Gly Leu Lys Gly Leu Val Pro Gly Asn 465 470 475 63478PRTAspergillus niger 63Met Ser Gly Leu Arg Phe Leu Asp Leu Ile Lys Pro Phe Thr Pro Leu 1 5 10 15 Leu Pro Glu Val Ala Ala Pro Glu Thr Lys Val Pro Phe Asn Gln Lys 20 25 30 Leu Met Trp Thr Gly Leu Thr Leu Leu Ile Phe Leu Val Met Ser Gln 35 40 45 Met Pro Leu Tyr Gly Ile Val Ser Ser Asp Thr Ser Asp Pro Leu Tyr 50 55 60 Trp Leu Arg Met Met Leu Ala Ser Asn Arg Gly Thr Leu Met Glu Leu 65 70 75 80 Gly Ile Thr Pro Ile Ile Ser Ser Gly Met Val Phe Gln Leu Leu Ala 85 90 95 Gly Thr His Leu Ile Asp Val Asn Leu Asp Leu Lys Thr Asp Arg Glu 100 105 110 Leu Tyr Gln Thr Ala Gln Lys Leu Phe Ala Ile Ile Leu Ser Phe Gly 115 120 125 Gln Ala Cys Val Tyr Val Leu Thr Gly Leu Tyr Gly Gln Pro Ser Asp 130 135 140 Leu Gly Ala Gly Ile Cys Val Leu Leu Ile Val Gln Leu Val Val Ala 145 150 155 160 Gly Leu Val Val Ile Leu Leu Asp Glu Leu Leu Gln Lys Gly Tyr Gly 165 170 175 Leu Gly Ser Gly Ile Ser Leu Phe Ile Ala Thr Asn Ile Cys Glu Ser 180 185 190 Ile Val Trp Lys Ala Phe Ser Pro Thr Thr Ile Asn Thr Gly Arg Gly 195 200 205 Pro Glu Phe Glu Gly Ala Ile Ile Ala Leu Phe His Leu Leu Leu Thr 210 215 220 Trp Ser Asp Lys Gln Arg Ala Leu Arg Glu Ala Phe Tyr Arg Gln Asn 225 230 235 240 Leu Pro Asn Ile Met Asn Leu Leu Ala Thr Leu Leu Val Phe Ala Ala 245 250 255 Val Ile Tyr Leu Gln Gly Phe Arg Val Glu Ile Pro Val Lys Ser Ser 260 265 270 Arg Gln Arg Gly Met Arg Gly Ser Tyr Pro Val Arg Leu Phe Tyr Thr 275 280 285 Ser Asn Met Pro Ile Met Leu Gln Ser Ala Leu Cys Ser Asn Ile Phe 290 295 300 Leu Ile Ser Gln Met Leu Tyr Ser Arg Phe Ser Asp Asn Leu Leu Val 305 310 315 320 Lys Leu Leu Gly Val Trp Glu Pro Arg Glu Gly Ser Ala Gln Leu His 325

330 335 Ala Ala Ser Gly Ile Ala Tyr Tyr Met Ser Pro Pro Leu Asn Phe Lys 340 345 350 Glu Ala Leu Leu Asp Pro Ile His Thr Ala Val Tyr Ile Thr Phe Met 355 360 365 Leu Val Ala Cys Ala Leu Phe Trp Lys Thr Trp Ile Glu Val Ser Gly 370 375 380 Ser Ala Pro Arg Asp Val Ala Lys Gln Leu Lys Asp Gln Gly Leu Val 385 390 395 400 Met Ala Gly His Arg Glu Gln Ser Met Tyr Lys Glu Leu Lys Arg Val 405 410 415 Ile Pro Thr Ala Ala Ala Phe Gly Gly Ala Cys Ile Gly Ala Leu Ser 420 425 430 Val Ala Ser Asp Leu Leu Gly Ala Leu Gly Ser Gly Thr Gly Ile Leu 435 440 445 Leu Ala Val Thr Ile Ile Tyr Gly Tyr Phe Glu Ile Ala Ala Arg Glu 450 455 460 Gly Asp Ile Gly Ser Gly Leu Lys Gly Leu Val Pro Gly Asn 465 470 475

Claims

1. Method to improve the secretion of a protein of interest by a filamentous fungal cell comprising inducing a phenotype in the cell selected from the group consisting of: (i) a lowered ERAD, (ii) an elevated UPR that does not induce an elevated ERAD, (iii) an elevated UPR that does not induce an elevated ERAD, wherein ERAD is lowered

2. Method to improve the secretion of a protein of interest by a filamentous fungal cell, wherein the expression of a DNA sequence is modulated, said DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, or a homologue thereof.

3. Method according to claim 2, wherein the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, or a homologue thereof, is modulated, preferably up regulated.

4. Method according to claim 2 or 3, wherein the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or a homologue thereof, is modulated, preferably down regulated.

5. Method according to claims 2 to 4, characterized in that the expression level of at least one of the DNA sequences having the following SEQ ID NO or homologues thereof given below or a combination of at least one taken from each subgroup a), b) c), d), or e) given below, or a combination thereof is up regulated: a) 4, 25, 34, 40, b) 25, c) 10, 13, 22, 25, 28, 31, 31, d) 25, e) 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, and/or wherein the expression level of at least one of the DNA sequences selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or homologues thereof, or a combination thereof is down regulated.

6. Method according to claims 1 to 5, wherein expression of the native sec61 gene is preferably impaired, said method further comprising production in the filamentous fungal cell of: (i) a polypeptide according to SEQ ID NO: 63, and/or (ii) a polypeptide according to SEQ ID NO: 63, wherein the amino acid at position 376 is replaced by phenylalanine, tyrosine or histidine.

7. Method according to any one of claims 2 to 6, characterized in that the expression level of a DNA sequence which is down regulated is lower in the obtained filamentous fungus than the expression level of the corresponding DNA sequence in the parental filamentous fungus the filamentous fungus originates from, preferably three times lower, more preferably four times lower, most preferably more than four times lower and even most preferably not detectable using northern, or western blotting or array technique.

8. Method according to any one of claims 2 to 7, characterized in that the expression level of a DNA sequence which is up regulated is higher in the obtained filamentous fungus than the expression level of the corresponding DNA sequence in the parental filamentous fungus the filamentous fungus originates from, preferably three times higher, more preferably four times higher and most preferably more than four times higher using northern, or western blotting or array technique.

9. Method according to any one of claims 1 to 8, wherein the filamentous fungus is selected from the group consisting of the genera Aspergillus, Trichoderma and Penicillium.

10. Method according to claim 9, characterized in that the Aspergillus is selected from the species Aspergillus niger, Aspergillus oryzae, Aspergillus sojae and the Trichoderma is selected from the species Trichoderma reesei.

11. Polynucleotide comprising a DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, or a homologue thereof, or a degenerated DNA sequence obtainable there from.

12. Polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO's: 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, or a homologue thereof.

13. Filamentous fungus displaying a modulated expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, or a homologue thereof.

14. Filamentous fungus, wherein the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, or a homologue thereof, is modulated, preferably up regulated.

15. Filamentous fungus, wherein the expression of at least one DNA sequence selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or a homologue thereof, is modulated, preferably down regulated.

16. Filamentous fungus, wherein the expression level of at least one of the DNA sequences having the following SEQ ID NO or homologues thereof given below or a combination of at least one taken from each subgroup a), b) c), d), or e) given below, or a combination thereof is up regulated: a) 4, 25, 34, 40, b) 25, c) 10, 13, 22, 25, 28, 31, 31, d) 25, e) 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, and/or wherein the expression level of at least one of the DNA sequences selected from the group consisting of SEQ ID NO's: 43, 46, 49, 52, 55, 58, or homologues thereof, or a combination thereof is down regulated.

17. Filamentous fungus according to claims 14 to 18, wherein expression of the native sec61 gene is preferably impaired, said filamentous fungus further synthesising intracellularly: (i) a sec61 polypeptide according to SEQ ID NO: 63, and/or (ii) a sec61 polypeptide according to SEQ ID NO: 63, wherein the amino acid at position 376 is replaced by phenylalanine, tyrosine or histidine.

18. Filamentous fungus according to claims 13 to 17, transformed with the polynucleotide of claim 12.

19. Filamentous fungus according to claim 13 or 18, wherein the filamentous fungus further comprises a DNA sequence encoding a protein of interest.

20. Filamentous fungus according to claim 19, wherein the DNA sequence encoding a protein of interest is operably linked to a promoter and to a secretion signal.

21. Method for the production of a protein of interest comprising culturing the filamentous fungus of claim 19 or 20 under conditions conducive to expression of the protein and optionally recovering the produced protein.

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