IP Library Granted Patent US 12,234,493
Granted Patent B2
US 12,234,493 · App. 18/333,776 · Granted Feb 25, 2025

Compositions for saccharification of cellulosic material

Inventors: Brett McBrayer (Sacramento, CA); Elena Vlasenko (Davis, CA); Tarana Shaghasi (Guerneville, CA)
Assignee: Novozymes, Inc.
C12P19/14C12N1/22C12N9/2402C12N9/2437C12N9/2445C12N9/248C12P19/02C12P19/12C12P21/02C12Y302/01C12Y302/01004C12Y302/01008C12Y302/01021C12Y302/01091C12P2203/00C12Y302/01037Y02P20/52
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Quick Facts
Patent No.
US 12,234,493
App. No.
18/333,776
Granted
Feb 25, 2025
Kind
B2
Abstract

The present invention relates to enzyme compositions for high temperature saccharification of cellulosic material and to uses thereof.

Claims (69)

1. An isolated recombinant host cell encoding an enzyme composition comprising:

(I) a polypeptide having cellobiohydrolase I activity selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6;

(II) a polypeptide having beta-glucosidase activity; and

(III) a polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

2. The isolated recombinant host cell of claim 1 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

3. The isolated recombinant host cell of claim 1 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

4. The isolated recombinant host cell of claim 1 , wherein the enzyme composition further comprises a polypeptide having cellobiohydrolase II activity.

5. The isolated recombinant host cell of claim 4 , wherein the polypeptide having cellobiohydrolase II activity is from Talaromyces.

6. The isolated recombinant host cell of claim 1 , wherein the polypeptide having beta-glucosidase activity is from Talaromyces.

7. The isolated recombinant host cell of claim 1 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

8. The isolated recombinant host cell of claim 1 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

9. The isolated recombinant host cell of claim 1 , wherein the enzyme composition further comprises a polypeptide having endoglucanase II activity.

10. The isolated recombinant host cell of claim 1 , wherein the enzyme composition further comprises a GH61 polypeptide having cellulolytic enhancing activity.

11. The isolated recombinant host cell of claim 1 , wherein the enzyme composition further comprises a polypeptide having xylanase activity.

12. The isolated recombinant host cell of claim 11 , wherein the enzyme composition further comprises a Family 10 polypeptide having xylanase activity or a Family 11 polypeptide having xylanase activity.

13. The isolated recombinant host cell of claim 1 , wherein the enzyme composition further comprises a polypeptide having beta-xylosidase activity.

14. The isolated recombinant host cell of claim 13 , wherein the polypeptide having beta-xylosidase activity is a GH3 beta-xylosidase.

15. The isolated recombinant host cell of claim 13 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

16. The isolated recombinant host cell of claim 13 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

17. The isolated recombinant host cell of claim 13 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

18. A method of producing an enzyme composition, comprising: (a) cultivating the recombinant host cell of claim 1 under conditions conducive for production of the enzyme composition; and (b) recovering the enzyme composition.

19. A method for degrading or converting a cellulosic material, comprising: treating the cellulosic material with an enzyme composition comprising:

(I) a polypeptide having cellobiohydrolase I activity selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6;

(II) a polypeptide having beta-glucosidase activity; and

(III) a polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62;

wherein the degrading or converting of the cellulosic material is performed with the enzyme composition at a temperature in the range of about 40° C. to about 70° C.

20. The method of claim 19 , wherein saccharification of the cellulosic material is performed with the enzyme composition at a temperature in the range of about 50° C. to about 65° C.

21. The method of claim 19 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

22. The method of claim 19 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

23. The method of claim 19 , wherein the enzyme composition further comprises a polypeptide having cellobiohydrolase II activity.

24. The method of claim 23 , wherein the polypeptide having cellobiohydrolase II activity is from Talaromyces.

25. The method of claim 19 , wherein the polypeptide having beta-glucosidase activity is from Talaromyces.

26. The method of claim 19 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

27. The method of claim 19 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

28. The method of claim 19 , wherein the enzyme composition further comprises a polypeptide having endoglucanase II activity.

29. The method of claim 19 , wherein the enzyme composition further comprises a GH61 polypeptide having cellulolytic enhancing activity.

30. The method of claim 19 , wherein the enzyme composition further comprises a polypeptide having xylanase activity.

31. The method of claim 19 , wherein the enzyme composition further comprises a Family 10 polypeptide having xylanase activity or a Family 11 polypeptide having xylanase activity.

32. The method of claim 19 , wherein the enzyme composition further comprises a polypeptide having beta-xylosidase activity.

33. The method of claim 32 , wherein the polypeptide having beta-xylosidase activity is a GH3 beta-xylosidase.

34. The method of claim 32 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

35. The method of claim 32 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

36. The method of claim 32 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

37. A method for producing a fermentation product, comprising:

(a) saccharifying a cellulosic material with an enzyme composition comprising:

(I) a polypeptide having cellobiohydrolase I activity selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6;

(II) a polypeptide having beta-glucosidase activity; and

(III) a polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62;

wherein the saccharification of the cellulosic material is performed with the enzyme composition at a temperature in the range of about 40° C. to about 70° C.;

(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and

(c) recovering the fermentation product from the fermentation.

38. The method of claim 37 , wherein saccharification of the cellulosic material is performed with the enzyme composition at a temperature in the range of about 50° C. to about 65° C.

39. The method of claim 37 , wherein saccharification of the cellulosic material is performed with the enzyme composition at a temperature in the range of about 50° C. to about 65° C.

40. The method of claim 37 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

41. The method of claim 37 , wherein the polypeptide having cellobiohydrolase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 158; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 157, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 157, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 6.

42. The method of claim 37 , wherein the composition further comprises a polypeptide having cellobiohydrolase II activity.

43. The method of claim 42 , wherein the polypeptide having cellobiohydrolase II activity is from Talaromyces.

44. The method of claim 37 , wherein the polypeptide having beta-glucosidase activity is from Talaromyces.

45. The method of claim 37 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

46. The method of claim 37 , wherein the polypeptide having endoglucanase I activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 62; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 61, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 61, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 61; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 62.

47. The method of claim 37 , wherein the enzyme composition further comprises a polypeptide having endoglucanase II activity.

48. The method of claim 37 , wherein the enzyme composition further comprises a GH61 polypeptide having cellulolytic enhancing activity.

49. The method of claim 37 , wherein the enzyme composition further comprises a polypeptide having xylanase activity.

50. The method of claim 37 , wherein the enzyme composition further comprises a Family 10 polypeptide having xylanase activity or a Family 11 polypeptide having xylanase activity.

51. The method of claim 37 , wherein the enzyme composition further comprises a polypeptide having beta-xylosidase activity.

52. The method of claim 51 , wherein the polypeptide having beta-xylosidase activity is a GH3 beta-xylosidase.

53. The method of claim 51 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

54. The method of claim 51 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

55. The method of claim 51 , wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 206; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 205, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 205, or (iii) the full-length complement of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 205; and (d) a polypeptide comprising the mature polypeptide of SEQ ID NO: 206.

Assignments (1)
CONFIRMATORY LICENSE Recorded Feb 3, 2025
From: NOVOZYMES,INC.
To: UNITED STATES DEPARTMENT OF ENERGY
Reel/Frame 070083/0846 →
Continuity (9)
Continuation 17372667 · Jul 12, 2021
Continuation 16840012 · Apr 3, 2020
Division 16115930 · Aug 29, 2018
Division 15422245 · Feb 1, 2017
Division 14885555 · Oct 16, 2015
Division 14052360 · Oct 11, 2013
Division 12940952 · Nov 5, 2010
Provisional Application 61259014 · Nov 6, 2009
Related Publication 20240018560A1 · Jan 18, 2024
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