Methods for enhancing the degradation or conversion of cellulosic material

Patent metadata

Field	Value
Publication number	US-9624518-B2
Application number	US-201214112023-A
Country	US
Kind code	B2
Filing date	Apr 27, 2012
Priority date	Apr 29, 2011
Publication date	Apr 18, 2017
Grant date	Apr 18, 2017

Abstract

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.

First claim

What is claimed is:

1. A method for degrading or converting a cellulosic material, comprising: treating the cellulosic material under high temperature conditions with a cellulolytic enzyme composition comprising 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; and wherein the high temperature conditions are in the range of about 54° C. to about 70° C.

2. The method of claim 1 , wherein the GH61 polypeptide comprises SEQ ID NO: 2 or the mature polypeptide thereof.

3. 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.

4. The method of claim 1 , further comprising recovering the degraded or converted cellulosic material.

5. The method of claim 4 , wherein the degraded or converted cellulosic material is a sugar.

6. The method of claim 1 , wherein the cellulosic material is pretreated.

7. The method of claim 3 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.

8. The method of claim 3 , 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.

9. The method of claim 5 , wherein the sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.

10. 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.

11. The method of claim 1 , wherein the GH61 polypeptide has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.

12. The method of claim 1 , wherein the GH61 polypeptide has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.

13. The method of claim 1 , wherein the GH61 polypeptide has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.

14. The method of claim 1 , wherein the GH61 polypeptide has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.

15. The method of claim 1 , wherein the GH61 polypeptide consists of the mature polypeptide of SEQ ID NO: 2.

16. 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.

17. 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.

18. 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.

19. 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.

20. 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.

21. 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.

22. 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.

Assignees

• Shaghasi Tarana
• Mcbrayer Brett
• Novozymes Inc

Inventors

• Shaghasi Tarana
• Mcbrayer Brett

Classifications

• C07K14/37

  from fungi · CPC title

• C12P19/02 Primary

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• C12P19/14 Primary

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C11D3/386

  Preparations containing enzymes {, e.g. protease or amylase} · CPC title

• C11D3/3869 Primary

  Enzyme enhancers or mediators · CPC title