Polypeptides having cellulolytic enhancing activity and nucleic acids 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 for producing and using the polypeptides.
1. A method for degrading a cellulosic material, comprising: treating the cellulosic material with a composition comprising one or more cellulolytic proteins and a GH61polypeptide having cellulolytic enhancing activity, wherein the composition comprising the GH61polypeptide having cellulolytic enhancing activity increases the degradation of cellulosic material compared to a composition without the GH61 polypeptide having cellulolytic enhancing activity, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide comprising an amino acid sequence having at least 90sequence identity to amino acids 19 to 226 of SEQ ID NO: 8;
(b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, wherein 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, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; and
(c) a GH61 polypeptide encoded by a polynucleotide having at least 90% sequence identity to nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7.
2. The method of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 90% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
3. The method of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 95% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
4. The method of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 97% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
5. The method of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 98% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
6. The method of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 99% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
7. The method of claim 1 , wherein the GH61 polypeptide comprises SEQ ID NO: 8 or amino acids 19 to 226 of SEQ ID NO: 8.
8. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, wherein 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, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.
9. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, wherein very 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, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.
10. The method of claim 1 , wherein the GH61 polypeptide is encoded by the polynucleotide contained in plasmid pTter61E which is contained in E. coli NRRL B-30814.
11. The method of claim 1 , wherein the cellulosic material is selected from the group consisting of herbaceous material, agricultural residue, forestry residue, municipal solid waste, waste paper, and pulp and paper mill residue.
12. The method of claim 1 , wherein the cellulosic material is corn stover.
13. The method of claim 1 , wherein the one or more cellulolytic proteins are selected from the group consisting of a cellulase, endoglucanase, cellobiohydrolase, and beta-glucosidase.
14. The method of claim 1 , further comprising treating the cellulosic material with one or more enzymes selected from the group consisting of a hemicellulase, esterase, protease, laccase, peroxidase, or a mixture thereof.
15. The method of claim 1 , wherein the method is a step in a simultaneous saccharification and fermentation process (SSF).
16. The method of claim 1 , further comprising recovering the degraded cellulosic material.
17. The method of claim 16 , wherein the degraded cellulosic material is a sugar.
18. The method of claim 17 , wherein the sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.
19. The method of claim 1 , wherein the cellulolytic protein and/or the GH61 polypeptide having cellulolytic enhancing activity are in the form of a fermentation broth with or without cells.