IP Library Granted Patent US 10,577,622
Granted Patent B2
US 10,577,622 · App. 16/115,949 · Granted Mar 3, 2020

Polypeptides having beta-xylosidase activity and polynucleotides encoding same

Inventors: Yu Zhang (Beijing, CN); Ye Liu (Beijing, CN); Junxin Duan (Beijing, CN); Lan Tang (Beijing, CN); Brett McBrayer (Sacramento, CA)
Assignee: Novozymes Inc.
C12N15/8242C12N9/248C12N9/2445C12N15/1137C12N15/8218C12N15/8246C12P7/10C12P19/02C12P19/14C12Y302/01008C12Y302/01021C12Y302/01037C12N2310/14C12N2310/141C12P2203/00Y02E50/17Y02E50/343Y02P20/52
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Quick Facts
Patent No.
US 10,577,622
App. No.
16/115,949
Granted
Mar 3, 2020
Kind
B2
Abstract

The present invention relates to isolated polypeptides having beta-xylosidase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Claims (59)

1. A process for degrading or converting a cellulosic or xylan-containing material, comprising: treating the cellulosic or xylan-containing material with an enzyme composition comprising a polypeptide having beta-xylosidase activity, wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of:

(a) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10;

(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 9, wherein very 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 70° C.;

(c) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 9 or the cDNA sequence thereof;

(d) a variant of the mature polypeptide of SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at one or more positions and having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10; and

(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has beta-xylosidase activity.

2. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 10.

3. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 10.

4. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 10.

5. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 10.

6. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 10.

7. The process of claim 1 , wherein the polypeptide having beta-xylosidase activity comprises SEQ ID NO: 10 or the mature polypeptide of SEQ ID NO: 10.

8. The process of claim 1 , wherein the cellulosic or xylan-containing material is pretreated.

9. The process of claim 1 , wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

10. The process of claim 9 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.

11. The process of claim 9 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.

12. The process of claim 1 , further comprising recovering the degraded or converted cellulosic or xylan-containing material.

13. The process of claim 12 , wherein the degraded or converted cellulosic or xylan-containing material is a sugar.

14. The process of claim 13 , wherein the sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.

15. A process for producing a fermentation product, comprising:

(a) saccharifying a cellulosic or xylan-containing material with an enzyme composition comprising a polypeptide having beta-xylosidase activity, wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of:

(i) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10;

(ii) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 9, wherein very 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 70° C.;

(iii) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 9 or the cDNA sequence thereof;

(iv) a variant of the mature polypeptide of SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at one or more positions and having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10; and

(v) a fragment of the polypeptide of (i), (ii), (iii), or (iv) that has beta-xylosidase activity;

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

(c) recovering the fermentation product from the fermentation.

16. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 10.

17. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 10.

18. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 10.

19. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 10.

20. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 10.

21. The process of claim 15 , wherein the polypeptide having beta-xylosidase activity comprises SEQ ID NO: 10 or the mature polypeptide of SEQ ID NO: 10.

22. The process of claim 15 , wherein the cellulosic or xylan-containing material is pretreated.

23. The process of claim 15 , wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

24. The process of claim 23 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.

25. The process of claim 23 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.

26. The process of claim 15 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.

27. The process of claim 15 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.

28. A process of fermenting a cellulosic or xylan-containing material, comprising: fermenting the cellulosic or xylan-containing material with one or more fermenting microorganisms, wherein the cellulosic or xylan-containing material is saccharified with an enzyme composition comprising a polypeptide having beta-xylosidase activity, wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of:

(a) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10;

(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 9, wherein very 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 70° C.;

(c) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 9 or the cDNA sequence thereof;

(d) a variant of the mature polypeptide of SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at one or more positions and having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 10; and

(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has beta-xylosidase activity.

29. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 10.

30. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 10.

31. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 10.

32. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 10.

33. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 10.

34. The process of claim 28 , wherein the polypeptide having beta-xylosidase activity comprises SEQ ID NO: 10 or the mature polypeptide of SEQ ID NO: 10.

35. The process of claim 28 , wherein the cellulosic or xylan-containing material is pretreated.

36. The process of claim 28 , wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

37. The process of claim 36 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.

38. The process of claim 36 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.

39. The process of claim 28 , wherein the fermenting of the cellulosic or xylan-containing material produces a fermentation product.

40. The process of claim 39 , further comprising recovering the fermentation product from the fermentation.

41. The process of claim 40 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.

Assignments (1)
CONFIRMATORY LICENSE Recorded Feb 6, 2025
From: NOVOZYMES,INC.
To: UNITED STATES DEPARTMENT OF ENERGY
Reel/Frame 070126/0714 →
Priority Claims (1)
CN PCT/CN2011/082627 · Nov 22, 2011 · national
Continuity (4)
Division 15621771 · Jun 13, 2017
Division 14356482
Provisional Application 61569910 · Dec 13, 2011
Related Publication 20180371482A1 · Dec 27, 2018