Methods for enhancing the degradation or conversion of cellulosic material
The present invention relates to methods for degrading or converting a cellulosic material and for producing substances from the cellulosic material under high temperature conditions.
1. A method of fermenting a cellulosic material, comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified under high temperature conditions with an enzyme composition in the presence of a GH61 polypeptide having cellulolytic enhancing activity selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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.;
(c) a GH61 polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof; and
(d) a fragment of the GH61 polypeptide of (a), (b), or (c) that has cellulolytic enhancing activity;
wherein the high temperature conditions are a temperature of about 54° C. to about 70° C. for about 6 to about 168 hours at a pH of about 3 to about 8 and a dry solids content of a cellulosic material of about 5 to about 50 wt %; wherein an effective amount of the GH61 polypeptide is about 0.01 to about 50.0 mg per g of the cellulosic material.
2. The method of claim 1 , wherein the GH61 polypeptide has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
3. The method of claim 1 , wherein the GH61 polypeptide has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
4. The method of claim 1 , wherein the GH61 polypeptide has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
5. The method of claim 1 , wherein the GH61 polypeptide has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
6. The method of claim 1 , wherein the GH61 polypeptide comprises SEQ ID NO: 2 or the mature polypeptide thereof.
7. The method of claim 1 , wherein the GH61 polypeptide consists of the mature polypeptide of SEQ ID NO: 2.
8. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, 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 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.
9. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide having at least 96% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
10. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide having at least 97% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
11. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
12. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide having at least 99% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
13. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide comprising the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
14. The method of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide consisting of the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof.
15. The method of claim 1 , wherein the cellulolytic enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
16. The method of claim 15 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.
17. The method of claim 15 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.
18. The method of claim 1 , wherein the cellulosic material is pretreated.
19. The method of claim 1 , wherein the fermenting of the cellulosic material produces a fermentation product.
20. The method of claim 19 , further comprising recovering the fermentation product from the fermentation.
21. The method of claim 20 , wherein the fermentation product is an alcohol, an organic acid, a ketone, an amino acid, an alkane, a cycloalkane, an alkene, isoprene, polyketide, or a gas.
22. The method of claim 1 , wherein the cellulolytic enzyme composition and/or the GH61 polypeptide having cellulolytic enhancing activity are in the form of a fermentation broth with or without cells.