Beta-glucosidase variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9688975-B2
Application number	US-201113876802-A
Country	US
Kind code	B2
Filing date	Sep 30, 2011
Priority date	Oct 1, 2010
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

The present invention relates to beta-glucosidase variants, e.g. beta-glucosidase variants of a parent Family GH3A beta-glucosidase from Aspergillus fumigatus . The present invention also relates to polynucleotides encoding the beta-glucosidase variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the beta-glucosidase variants.

First claim

What is claimed is:

1. A beta-glucosidase variant, comprising a substitution to a parent beta-glucosidase at one or more positions corresponding to positions selected from the group consisting of 89, 91, 100, 140, 186, 283, and 456 of SEQ ID NO: 2, wherein the variant has beta-glucosidase activity, and wherein the variant comprises an amino acid sequence having at least 95% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

2. The variant of claim 1 , which comprises one or more substitutions selected from the group consisting of L89M, G91L, F100D, I140V, I186V, S283G, and N456E.

3. The variant of claim 1 , which comprises at least one of the substitutions L89M, G91L, F100D, I140V, I186V, S283G, or N456E, wherein the variant comprises an amino acid sequence having at least 98% but less than 100% sequence identify to amino acids 20 to 863 of SEQ ID NO: 2.

4. The variant of claim 3 , wherein the variant comprises an amino acid sequence having at least 99% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

5. An isolated polynucleotide encoding the variant of claim 1 .

6. A host cell comprising an expression vector, wherein the expression vector comprises the polynucleotide of claim 5 .

7. A method of producing a variant, said method comprising:
(a) cultivating the host cell of claim 6 under conditions suitable for the expression of the variant; and
(b) recovering the variant.

8. A method for degrading a cellulosic material, said method comprising: treating the cellulosic material with an enzyme composition, wherein the composition comprises the variant of claim 1 .

9. The method of claim 8 , said method further comprising recovering the degraded cellulosic material.

10. The method of claim 8 , wherein the enzyme composition comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an expansin, an esterase, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

11. A method for producing a fermentation product, said method comprising:
(a) saccharifying a cellulosic material with an enzyme composition, wherein the composition comprises the variant of claim 1 ;
(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.

12. A whole broth formulation or cell culture composition comprising the variant of claim 1 .

13. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having at least 95% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

14. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having at least 97% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

15. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having at least 98% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

16. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having at least 99% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

17. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having amino acids 20 to 863 of SEQ ID NO: 2.

18. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 89 of SEQ ID NO: 2.

19. The variant of claim 18 , wherein the substitution is to Met.

20. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 91 of SEQ ID NO: 2.

21. The variant of claim 20 , wherein the substitution is to Leu.

22. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 100 of SEQ ID NO: 2.

23. The variant of claim 22 , wherein the substitution is to Asp.

24. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 140 of SEQ ID NO: 2.

25. The variant of claim 24 , wherein the substitution is to Val.

26. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 186 of SEQ ID NO: 2.

27. The variant of claim 26 , wherein the substitution is to Val.

28. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 283 of SEQ ID NO: 2.

29. The variant of claim 26 , wherein the substitution is to Gly.

30. The variant of claim 1 , which comprises a substitution to a parent beta-glucosidase at a position corresponding to position 456 of SEQ ID NO: 2.

31. The variant of claim 30 , wherein the substitution is to Glu.

32. The variant of claim 1 , which further comprises a substitution to a parent beta-glucosidase at a position corresponding to position 512 of SEQ ID NO: 2.

33. The variant of claim 32 , wherein the substitution is to Tyr.

34. The variant of claim 1 , which comprises a Met at a position corresponding to position 89 of SEQ ID NO: 2, a Leu at a position corresponding to position 91 of SEQ ID NO. 2, a Val at a position corresponding to position 140 of SEQ ID NO: 2, and a Val at a position corresponding to position 186 of SEQ ID NO: 2.

35. The variant of claim 1 , which comprises an Asp at a position corresponding to position 100 of SEQ ID NO: 2, a Gly at a position corresponding to position 283 of SEQ ID NO: 2, and a Glu at a position corresponding to position 456 of SEQ ID NO: 2.

36. The variant of claim 1 , wherein the variant further comprises a substitution corresponding to position 512 of SEQ ID NO: 2, wherein the variant has beta-glucosidase activity, and wherein the variant comprises an amino acid sequence having at least 98% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

37. The variant of claim 36 , wherein the variant comprises an amino acid sequence having at least 99% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

38. The variant of claim 1 , wherein the parent beta-glucosidase comprises an amino acid sequence having at least 96% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

39. The variant of claim 1 , wherein the variant comprises an amino acid sequence having at least 96% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

40. The variant of claim 1 , wherein the variant comprises an amino acid sequence having at least 97% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

41. The variant of claim 1 , wherein the variant comprises an amino acid sequence having at least 98% but less than 100% sequence identity to amino acids 20 to 863 of SEQ ID NO: 2.

42. The variant of claim 1 , wherein the variant comprises an amino acid sequence having at least 99% but less than 100% sequence identity to amin

Assignees

• Wogulis Mark
• Harris Paul
• Osborn David
• Novozymes Inc

Inventors

• Wogulis Mark
• Harris Paul
• Osborn David

Classifications

• C12Y302/01021

  Beta-glucosidase (3.2.1.21) · CPC title

• C12P7/54

  Acetic acid (vinegar C12J) · CPC title

• C12P7/44

  Polycarboxylic acids · CPC title

• C12P13/14

  Glutamic acid; Glutamine · CPC title

• C12P13/04

  Alpha- or beta- amino acids {(other amino acids C12P13/005)} · CPC title