Polypeptides having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-10227614-B2
Application number	US-201715400361-A
Country	US
Kind code	B2
Filing date	Jan 6, 2017
Priority date	Nov 18, 2011
Publication date	Mar 12, 2019
Grant date	Mar 12, 2019

Abstract

The present invention relates to isolated polypeptides having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity and isolated 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.

First claim

The invention claimed is:

1. A nucleic acid construct comprising an isolated polynucleotide encoding a polypeptide having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polypeptide in a recombinant host cell, and wherein the polypeptide having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity, is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8;
(b) a polypeptide comprising amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8; and
(c) a fragment of the polypeptide of amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8, wherein the fragment has beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity.

2. A recombinant host cell transformed with the nucleic acid construct of claim 1 .

3. A method of producing a polypeptide having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity, comprising:
(a) cultivating the recombinant host cell of claim 2 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.

4. A transgenic plant, plant part or plant cell transformed with the nucleic acid construct of claim 1 .

5. A method of producing a polypeptide having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity, comprising:
(a) cultivating the transgenic plant or plant cell of claim 4 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.

6. A process of fermenting a cellulosic material or xylan-containing material, comprising: fermenting the cellulosic material or xylan-containing material with one or more fermenting microorganisms, wherein the cellulosic material or xylan-containing material is saccharified with an enzyme composition comprising a polypeptide having beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity, wherein the polypeptide is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8;
(b) a polypeptide comprising amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8; and
(c) a fragment of the polypeptide of amino acids 23 to 950 of SEQ ID NO: 2, amino acids 23 to 868 of SEQ ID NO: 4, amino acids 20 to 860 of SEQ ID NO: 6, or amino acids 20 to 737 of SEQ ID NO: 8, wherein the fragment has beta-glucosidase activity, beta-xylosidase activity, or beta-glucosidase and beta-xylosidase activity.

7. The process of claim 6 , wherein the fermenting of the cellulosic material or xylan-containing material produces a fermentation product.

8. The process of claim 7 , further comprising recovering the fermentation product from the fermentation.

9. The method of claim 3 , wherein the polypeptide has at least 90% sequence identity to amino acids 23 to 950 of SEQ ID NO: 2.

10. The method of claim 3 , wherein the polypeptide has at least 95% sequence identity to amino acids 23 to 950 of SEQ ID NO: 2.

11. The method of claim 3 , wherein the polypeptide has at least 97% sequence identity to amino acids 23 to 950 of SEQ ID NO: 2.

12. The method of claim 3 , wherein the polypeptide comprises amino acids 23 to 950 of SEQ ID NO: 2.

13. The method of claim 3 , wherein the polypeptide has at least 90% sequence identity to amino acids 23 to 868 of SEQ ID NO: 4.

14. The method of claim 3 , wherein the polypeptide has at least 95% sequence identity to amino acids 23 to 868 of SEQ ID NO: 4.

15. The method of claim 3 , wherein the polypeptide has at least 97% sequence identity to amino acids 23 to 868 of SEQ ID NO: 4.

16. The method of claim 3 , wherein the polypeptide comprises amino acids 23 to 868 of SEQ ID NO: 4.

17. The method of claim 3 , wherein the polypeptide has at least 90% sequence identity to amino acids 20 to 860 of SEQ ID NO: 6.

18. The method of claim 3 , wherein the polypeptide has at least 95% sequence identity to amino acids 20 to 860 of SEQ ID NO: 6.

19. The method of claim 3 , wherein the polypeptide has at least 97% sequence identity to amino acids 20 to 860 of SEQ ID NO: 6.

20. The method of claim 3 , wherein the polypeptide comprises amino acids 20 to 860 of SEQ ID NO: 6.

21. The method of claim 3 , wherein the polypeptide has at least 90% sequence identity to amino acids 20 to 737 of SEQ ID NO: 8.

22. The method of claim 3 , wherein the polypeptide has at least 95% sequence identity to amino acids 20 to 737 of SEQ ID NO: 8.

23. The method of claim 3 , wherein the polypeptide has at least 97% sequence identity to amino acids 20 to 737 of SEQ ID NO: 8.

24. The method of claim 3 , wherein the polypeptide comprises amino acids 20 to 737 of SEQ ID NO: 8.

25. The process of claim 6 , wherein the cellulosic material or xylan-containing material is pretreated.

26. The process of claim 6 , wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a GH61 polypeptide, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

27. The process of claim 7 , wherein the fermentation product is an alcohol, an alkane, an alkene, an amino acid, a cycloalkane, a gas, isoprene, a ketone, an organic acid, or polyketide.

Assignees

• Novozymes As
• Novozymes Inc

Inventors

• Morant Marc

Classifications

• Y02E50/16

  Cross-Sectional Technologies · mapped topic

• C12Y302/01008

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title

• C12P2203/00

  Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source · CPC title

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12N9/2437 Primary

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title