Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having cellulolytic enhancing 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.
1. An isolated GH61 polypeptide having cellulolytic enhancing activity, selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18to 246 of SEQ ID NO: 2;
(b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) nucleotides 52 to 875 of SEQ ID NO: 1, (ii) the cDNA sequence contained in the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) the full-length complement of (i) or (ii), wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5X 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 2X SSC, 0.2% SDS at 70° C.;
(c) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 875 of SEQ ID NO: 1 or the cDNA sequence thereof; and
(d) an isolated fragment of a GH61 polypeptide of (a), (b), or (c) that has cellulolytic enhancing activity.
2. The GH61 polypeptide of claim 1 , comprising amino acids 18 to 246 of SEQ ID NO: 2.
3. The GH61 polypeptide of claim 1 , which is encoded by the polynucleotide contained in pSMai216 which is contained in E. coli NRRL B-50301 deposited with the Northern Regional Research Center of the Agriculture Research Service Patent Culture Collection.
4. A nucleic acid construct comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 1 operably linked to one or more heteroloqous control sequences that direct the production of the polypeptide in an expression host.
5. A method of producing the polypeptide of claim 1 , said method comprising:
(a) cultivating an isolated cell, which in its wild-type form produces the polypeptide, under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
6. A method of producing a GH61 polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating a recombinant host cell comprising the polynucleotide of claim 4 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
7. A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding the polypeptide of claim 1 , wherein the polypeptide comprises a signal peptide directing the polypeptide into the secretory pathway.
8. A method of producing a GH61 polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating the transgenic plant or plant cell of claim 7 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
9. A DNA construct comprising a nucleic acid encoding the signal peptide comprising or consisting of amino acids 1 to 17 of SEQ ID NO: 2operably linked to a heterologous nucleic acid.
10. A method of producing a protein, said method comprising:
(a) cultivating a recombinant host cell comprising the construct of claim 9 under conditions conducive for production of the protein; and
(b) recovering the protein.
11. A method for degrading or converting a cellulosic material comprising treating the cellulosic material with an enzyme composition in the presence of the GH61 polypeptide having cellulolytic enhancing activity of claim 1 .
12. The method of claim 11 , said method further comprising recovering the degraded cellulosic material.
13. A method for producing a fermentation product, said method comprising:
(a) saccharifying a cellulosic material with an enzyme composition in the presence of the GH61 polypeptide having cellulolytic enhancing activity 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.
14. A method 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 polypeptide having cellulolytic enhancing activity of claim 1 .
15. The method of claim 14 , wherein the fermenting of the cellulosic material produces a fermentation product.
16. The method of claim 15 , said method further comprising recovering the fermentation product from the fermentation.
17. A detergent composition comprising the polypeptide of claim 1 and a surfactant.
18. The GH61 polypeptide of claim 1 , having at least 95% sequence identity to amino acids 18 to 246 of SEQ ID NO: 2.
19. The GH61 polypeptide of claim 18 , having at least 96% sequence identity to amino acids 18 to 246 of SEQ ID NO: 2.
20. The GH61 polypeptide of claim 19 , having at least 97% sequence identity to amino acids 18 to 246 of SEQ ID NO: 2.
21. The GH61 polypeptide of claim 20 , having at least 98% sequence identity to amino acids 18 to 246 of SEQ ID NO: 2.
22. The GH61 polypeptide of claim 21 , having at least 99% sequence identity to amino acids 18 to 246 of SEQ ID NO: 2.
23. The GH61 polypeptide of claim 1 , which is encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 875 of SEQ ID NO: 1 or the cDNA sequence thereof.
24. The GH61 polypeptide of claim 1 , which consists of amino acids 18 to 246 of SEQ ID NO: 2 or a fragment thereof having cellulolytic enhancing activity.
25. The GH61 polypeptide of claim 1 , which is encoded by a polynucleotide comprising nucleotides 52 to 875 of SEQ ID NO: 1 or the cDNA sequence thereof.
26. The GH61 polypeptide of claim 1 , which is encoded by a polynucleotide consisting of nucleotides 52 to 875 of SEQ ID NO: 1 or the cDNA sequence thereof.
27. The method of claim 13 , wherein steps (a) and (b) are performed simultaneously.
28. An isolated GH61 polypeptide, comprising SEQ ID NO: 2.
29. A nucleic acid construct comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 28 operably linked to one or more heteroloqous control sequences that direct the production of the polypeptide in an expression host.
30. A recombinant host cell comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 28 .
31. A method of producing a GH61 polypeptide, said method comprising:
(a) cultivating the recombinant host cell of claim 30 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
32. A recombinant expression vector comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 1 .
33. A recombinant host cell comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 1 .
34. A recombinant expression vector comprising an isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 28 .