Hybrid polypeptides having cellobiohydrolase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9969994-B2
Application number	US-201615287332-A
Country	US
Kind code	B2
Filing date	Oct 6, 2016
Priority date	Dec 22, 2011
Publication date	May 15, 2018
Grant date	May 15, 2018

Abstract

The present invention relates to hybrid polypeptides having cellobiohydrolase activity. The present invention also relates to polynucleotides encoding the hybrid polypeptides; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and processes of using the hybrid polypeptides.

First claim

What is claimed is:

1. An isolated hybrid polypeptide having cellobiohydrolase activity, comprising:
(a) a first polypeptide fragment comprising a cellobiohydrolase catalytic domain at the N-terminal end of the hybrid polypeptide selected from the group consisting of (i) a polypeptide fragment having at least 96% sequence identity to amino acids 19 to 455 of SEQ ID NO: 2; (ii) a polypeptide fragment encoded by a polynucleotide having at least 96% sequence identity to nucleotides 55 to 1485 of SEQ ID NO: 1 or the cDNA sequence thereof; and (iii) a polypeptide fragment comprising or consisting of amino acids 19 to 455 of SEQ ID NO: 2 or a fragment thereof having cellobiohydrolase activity; and
(b) a second polypeptide fragment comprising a carbohydrate binding domain at the C-terminal end of the first polypeptide fragment selected from the group consisting of (i) a polypeptide fragment having at least 96% sequence identity to amino acids 485 to 529 of SEQ ID NO: 4; (ii) a polypeptide fragment encoded by a polynucleotide having at least 96% sequence identity to nucleotides 1453 to 1587 of SEQ ID NO: 3; and (iii) a polypeptide fragment comprising or consisting of amino acids 485 to 529 of SEQ ID NO: 4 or a fragment thereof having carbohydrate binding activity.

2. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment has at least 96% sequence identity to amino acids 19 to 455 of SEQ ID NO: 2.

3. The hybrid polypeptide of claim 1 , which further comprises a signal peptide at the N-terminal end of the first polypeptide fragment.

4. The hybrid polypeptide of claim 3 , wherein the signal peptide is the signal peptide of SEQ ID NO: 2.

5. The hybrid polypeptide of claim 4 , wherein the signal peptide is amino acids 1 to 21 of SEQ ID NO: 2.

6. An isolated polynucleotide encoding the hybrid polypeptide of claim 1 .

7. A nucleic acid construct comprising the polynucleotide of claim 6 .

8. An expression vector comprising the polynucleotide of claim 6 .

9. A host cell comprising the polynucleotide of claim 6 .

10. A method of producing a hybrid polypeptide having cellobiohydrolase activity, comprising:
(a) cultivating the host cell of claim 9 under conditions suitable for the expression of the hybrid polypeptide; and
(b) recovering the hybrid polypeptide.

11. A transgenic plant, plant part or plant cell transformed with the polynucleotide of claim 6 encoding a hybrid polypeptide.

12. A method of producing the hybrid polypeptide of claim 1 , comprising:
(a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding the hybrid polypeptide under conditions conducive for production of the hybrid polypeptide; and
(b) recovering the hybrid polypeptide.

13. A process for degrading or converting a cellulosic material, comprising:
treating the cellulosic material with an enzyme composition in the presence of the hybrid polypeptide of claim 1 .

14. The process of claim 13 , wherein the cellulosic material is pretreated.

15. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with an enzyme composition in the presence of the hybrid polypeptide 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.

16. The process of claim 15 , wherein the cellulosic material is pretreated.

17. A process of fermenting a cellulosic material, comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition in the presence of the hybrid polypeptide of claim 1 .

18. The process of claim 17 , wherein the cellulosic material is pretreated before saccharification.

19. A whole broth formulation or cell culture composition comprising the hybrid polypeptide of claim 1 .

20. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment has at least 97% sequence identity to amino acids 19 to 455 of SEQ ID NO: 2.

21. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment has at least 98% sequence identity to amino acids 19 to 455 of SEQ ID NO: 2.

22. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment has at least 99% sequence identity to amino acids 19 to 455 of SEQ ID NO: 2.

23. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment comprises amino acids 19 to 455 of SEQ ID NO: 2.

24. The hybrid polypeptide of claim 1 , wherein the first polypeptide fragment consists of amino acids 19 to 455 of SEQ ID NO: 2.

25. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment has at least 96% sequence identity to amino acids 485 to 529 of SEQ ID NO: 4.

26. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment has at least 97% sequence identity to amino acids 485 to 529 of SEQ ID NO: 4.

27. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment has at least 98% sequence identity to amino acids 485 to 529 of SEQ ID NO: 4.

28. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment has at least 99% sequence identity to amino acids 485 to 529 of SEQ ID NO: 4.

29. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment comprises amino acids 485 to 529 of SEQ ID NO: 4.

30. The hybrid polypeptide of claim 1 , wherein the second polypeptide fragment consists of amino acids 485 to 529 of SEQ ID NO: 4.

Assignees

• Novozymes Inc

Inventors

• Liu Ye
• Shaghasi Tarana

Classifications

• C07K2319/02

  containing a signal sequence · CPC title

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• C12P2203/00

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

• Y02E50/16

  Cross-Sectional Technologies · mapped topic

• C07K14/38

  from Aspergillus · CPC title