Methods and compositions for degrading cellulosic material

A method for producing a fermentation product, comprising:

• (a) saccharifying a cellulosic material with an enzyme composition comprising a CEL6 polypeptide having cellobiohydrolase II activity, a GH10 polypeptide having xylanase activity, and one or more cellulolytic enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase, wherein one or both of the CEL6 polypeptide having cellobiohydrolase II activity and the GH10 polypeptide having xylanase activity are foreign to the one or more cellulolytic enzymes;
• (b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms;
• and (c) recovering the organic fermentation product from the fermentation;
• wherein the CEL6 polypeptide having cellobiohydrolase II activity is selected from the group consisting of:
• (a) a polypeptide having cellobiohydrolase II activity comprising an amino acid sequence having a sequence identity of at least 90% to amino acids 18 to 482 of SEQ ID NO:
• 30;
• (b) a polypeptide having cellobiohydrolase II activity encoded by a polynucleotide that hybridizes under very high stringency conditions with nucleotides 52 to 1809 of SEQ ID NO:
• 29 or the full-length complement thereof, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.;
• (c) a polypeptide having cellobiohydrolase II activity encoded by a polynucleotide comprising a nucleotide sequence having a sequence identity to nucleotides 52 to 1809 of SEQ ID NO:
• 29 of at least 90%;
• and
• (d) a polypeptide having cellobiohydrolase II activity comprising amino acids 18 to 482 of SEQ ID NO:
• 30;
• and
• wherein the GH10 polypeptide having xylanase activity is selected from the group consisting of:
• (a) a polypeptide having xylanase activity comprising an amino acid sequence having a sequence identity of at least 90% to amino acids 20 to 369 of SEQ ID NO:
• 72, amino acids 19 to 414 of SEQ ID NO:
• 74, amino acids 18 to 364 of SEQ ID NO:
• 76, amino acids 20 to 323 of SEQ ID NO:
• 78, amino acids 20 to 397 of SEQ ID NO:
• 80, or amino acids 24 to 403 of SEQ ID NO:
• 99;
• (b) a polypeptide having xylanase activity encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of nucleotides 58 to 1107 of SEQ ID NO:
• 71, nucleotides 55 to 1242 of SEQ ID NO:
• 73, nucleotides 52 to 1145 of SEQ ID NO:
• 75, nucleotides 58 to 1400 of SEQ ID NO:
• 77, nucleotides 107 to 1415 of SEQ ID NO:
• 79, or nucleotides 70 to 1385 of SEQ ID NO:
• 98 or the full-length complement thereof, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.;
• (c) a polypeptide having xylanase activity encoded by a polynucleotide comprising a nucleotide sequence having a sequence identity of at least 90% to the mature polypeptide coding sequence of nucleotides 58 to 1107 of SEQ ID NO:
• 71, nucleotides 55 to 1242 of SEQ ID NO:
• 73, nucleotides 52 to 1145 of SEQ ID NO:
• 75, nucleotides 58 to 1400 of SEQ ID NO:
• 77, nucleotides 107 to 1415 of SEQ ID NO:
• 79, or nucleotides 70 to 1385 of SEQ ID NO:
• 98;
• and
• (d) a polypeptide having xylanase activity comprising amino acids 20 to 369 of SEQ ID NO:
• 72, amino acids 19 to 414 of SEQ ID NO:
• 74, amino acids 18 to 364 of SEQ ID NO:
• 76, amino acids 20 to 323 of SEQ ID NO:
• 78, amino acids 20 to 397 of SEQ ID NO:
• 80, or amino acids 24 to 403 of SEQ ID NO:
• 99;
• wherein the one or more cellulolytic enzymes comprise a beta-glucosidase, a Trichoderma reesei cellobiohydrolase I (CEL7A), a Trichoderma reesei cellobiohydrolase II (CEL6A), and a Trichoderma reesei endoglucanase I (CEL7B).

The method of claim 1, wherein the GH10 polypeptide having xylanase activity consists of amino acids 20 to 369 of SEQ ID NO:

• 72, amino acids 19 to 414 of SEQ ID NO:
• 74, amino acids 18 to 364 of SEQ ID NO:
• 76, amino acids 20 to 323 of SEQ ID NO:
• 78, amino acids 20 to 397 of SEQ ID NO:
• 80, or amino acids 24 to 403 of SEQ ID NO:
• 99.

A method of fermenting a cellulosic material, comprising:

• fermenting the cellulosic material with one or more fermenting microorganisms to produce a fermentation product, and optionally recovering the fermentation product, wherein the cellulosic material is saccharified with an enzyme composition comprising a CEL6 polypeptide having cellobiohydrolase II activity, a GH10 polypeptide having xylanase activity, and one or more cellulolytic enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase, wherein one or both of the CEL6 polypeptide having cellobiohydrolase II activity and the GH10 polypeptide having xylanase activity are foreign to the one or more cellulolytic enzymes;
• wherein the CEL6 polypeptide having cellobiohydrolase II activity is selected from the group consisting of:
• (a) a polypeptide having cellobiohydrolase II activity comprising an amino acid sequence having a sequence identity of at least 90% to amino acids 18 to 482 of SEQ ID NO:
• 30;
• (b) a polypeptide having cellobiohydrolase II activity encoded by a polynucleotide that hybridizes under very high stringency conditions with nucleotides 52 to 1809 of SEQ ID NO:
• 29 or the full-length complement thereof, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.;
• (c) a polypeptide having cellobiohydrolase II activity encoded by a polynucleotide comprising a nucleotide sequence having a sequence identity to nucleotides 52 to 1809 of SEQ ID NO:
• 29 of at least 90%;
• and
• (d) a polypeptide having cellobiohydrolase II activity comprising amino acids 18 to 482 of SEQ ID NO:
• 30;
• and
• wherein the GH10 polypeptide having xylanase activity is selected from the group consisting of:
• (a) a polypeptide having xylanase activity comprising an amino acid sequence having a sequence identity of at least 90% to amino acids 20 to 369 of SEQ ID NO:
• 72, amino acids 19 to 414 of SEQ ID NO:
• 74, amino acids 18 to 364 of SEQ ID NO:
• 76, amino acids 20 to 323 of SEQ ID NO:
• 78, amino acids 20 to 397 of SEQ ID NO:
• 80, or amino acids 24 to 403 of SEQ ID NO:
• 99;
• (b) a polypeptide having xylanase activity encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of nucleotides 58 to 1107 of SEQ ID NO:
• 71, nucleotides 55 to 1242 of SEQ ID NO:
• 73, nucleotides 52 to 1145 of SEQ ID NO:
• 75, nucleotides 58 to 1400 of SEQ ID NO:
• 77, nucleotides 107 to 1415 of SEQ ID NO:
• 79, or nucleotides 70 to 1385 of SEQ ID NO:
• 98 or the full-length complement thereof, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.;
• (c) a polypeptide having xylanase activity encoded by a polynucleotide comprising a nucleotide sequence having a sequence identity of at least 90% to the mature polypeptide coding sequence of nucleotides 58 to 1107 of SEQ ID NO:
• 71, nucleotides 55 to 1242 of SEQ ID NO:
• 73, nucleotides 52 to 1145 of SEQ ID NO:
• 75, nucleotides 58 to 1400 of SEQ ID NO:
• 77, nucleotides 107 to 1415 of SEQ ID NO:
• 79, or nucleotides 70 to 1385 of SEQ ID NO:
• 98;
• and
• (d) a polypeptide having xylanase activity comprising amino acids 20 to 369 of SEQ ID NO:
• 72, amino acids 19 to 414 of SEQ ID NO:
• 74, amino acids 18 to 364 of SEQ ID NO:
• 76, amino acids 20 to 323 of SEQ ID NO:
• 78, amino acids 20 to 397 of SEQ ID NO:
• 80, or amino acids 24 to 403 of SEQ ID NO:
• 99;
• wherein the one or more cellulolytic enzymes comprise a beta-glucosidase, a Trichoderma reesei cellobiohydrolase I (CEL7A), a Trichoderma reesei cellobiohydrolase II (CEL6A), and a Trichoderma reesei endoglucanase I (CEL7B).