Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having cellobiohydrolase 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.
1. An isolated polypeptide having cellobiohydrolase activity, selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the sequence of amino acids 26 to 532 of SEQ ID NO: 4;
(b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of the sequence of nucleotides 76 to 1596 of SEQ ID NO: 3 or the cDNA sequence thereof, wherein the 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, following standard Southern blotting procedures for 12 to 24 hours, followed by washing three times each for 15 minutes using 2× SSC, 0.2% SDS at 65° C.; and
(e) a fragment of the sequence of amino acids 26 to 532 of SEQ ID NO: 4 that has cellobiohydrolase activity.
2. The polypeptide of claim 1 , having at least 95% sequence identity to the sequence of amino acids 26to 532of SEQ ID NO: 4.
3. The polypeptide of claim 1 , having at least 97% sequence identity to the sequence of amino acids 26 to 532 of SEQ ID NO: 4.
4. The polypeptide of claim 1 , which is encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 3.
5. The polypeptide of claim 1 , which is a variant having at least 98% sequence identity to the sequence of amino acids 26 to 532 of SEQ ID NO: 4 comprising a substitution, deletion, and/or insertion at one or more positions.
6. The polypeptide of claim 1 , comprising the sequence of amino acids 26 to 532 of SEQ ID NO: 4.
7. The polypeptide of claim 1 , which is a fragment of the sequence of amino acids 26 to 532 of SEQ ID NO: 4, wherein the fragment has cellobiohydrolase activity.
8. A composition comprising a polypeptide of claim 1 , an endoglucanase, a GH61polypeptide, a beta-glucosidase, and a xylanase.
9. A composition comprising a polypeptide of claim 2 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
10. A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 1 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
11. A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 2 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
12. The process of claim 10 , further comprising recovering the degraded cellulosic material.
13. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 1 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase;
(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.
14. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 2 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase;
(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.
15. The process of claim 13 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.
16. The process of claim 13 , wherein the fermentation product is ethanol.
17. A recombinant host cell transformed with a nucleic acid construct or expression vector comprising a polynucleotide encoding the polypeptide of claim 1 , wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in the host cell.
18. A method of producing a polypeptide having cellobiohydrolase activity, comprising:
(a) cultivating the host cell of claim 17 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
19. An isolated polypeptide comprising a catalytic domain selected from the group consisting of:
(a) a catalytic domain having at least 90% sequence identity to the sequence of amino acids 26 to 459 of SEQ ID NO: 4; and
(b) a fragment of the sequence of amino acids 26to 459of SEQ ID NO: 4, which has cellobiohydrolase activity.
20. The polypeptide of claim 19 , which has at least 95% sequence identity to the sequence of amino acids 26 to 459 of SEQ ID NO: 4.
21. The polypeptide of claim 19 , wherein the catalytic domain comprises the sequence of amino acids 26 to 459 of SEQ ID NO: 4.
22. The polypeptide of claims 19 , further comprising a cellulose binding domain.
23. A composition comprising a polypeptide of claim 19 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
24. A composition comprising a polypeptide of claim 20 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
25. A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 19 , an endoglucanase, a GH61polypeptide, a beta-glucosidase, and a xylanase.
26. A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 20 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase.
27. The process of claim 25 , further comprising recovering the degraded cellulosic material.
28. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 19 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase;
(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.
29. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 20 , an endoglucanase, a GH61 polypeptide, a beta-glucosidase, and a xylanase;
(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.
30. The process of claim 28 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.
31. The process of claim 28 , wherein the fermentation product is ethanol.