IP Library Granted Patent US 8,148,495
Granted Patent B2
US 8,148,495 · App. 12/504,582 · Granted Apr 3, 2012

Methods for enhancing the degradation or conversion of cellulosic material

Assignee: Novozymes, Inc.
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Quick Facts
Patent No.
US 8,148,495
App. No.
12/504,582
Granted
Apr 3, 2012
Kind
B2
Abstract

The present invention relates to methods for degrading or converting a cellulosic material and for producing a substance from a cellulosic material.

Claims (21)

1. A method for producing a fermentation product, comprising:

(A) saccharifying a cellulosic material with an effective amount of one or more cellulolytic proteins in the presence of an effective amount of the polypeptide having cellulolytic enhancing activity, wherein the presence of the polypeptide having cellulolytic enhancing activity increases the degradation of cellulosic material compared to the absence of the polypeptide having cellulolytic enhancing activity and wherein the polypeptide having cellulolytic enhancing activity is selected from the group consisting of:

(i) a polypeptide comprising an amino acid sequence having at least 95% sequence identity with the mature polypeptide of SEQ ID NO: 2;

(ii) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1; and

(iii) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (a) the mature polypeptide coding sequence of SEQ ID NO: 1, (b) the genomic DNA sequence thereof, or (c) the full-length complement of (a) or (b), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/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.;

(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.

2. The method of claim 1 , wherein the one or more cellulolytic proteins are selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.

3. The method of claim 1 , further comprising treating the cellulosic material with an effective amount of one or more enzymes selected from the group consisting of a hemicellulase, an esterase, a protease, a laccase, a peroxidase, or a mixture thereof.

4. The method of claim 1 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.

5. The method of claim 1 , wherein the fermentation product is an alcohol, an organic acid, a ketone, an amino acid, or a gas.

6. The method of claim 1 , wherein the one or more cellulolytic proteins and/or the polypeptide having cellulolytic enhancing activity are in the form of a fermentation broth with or without cells.

7. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 95% sequence identity with the mature polypeptide of SEQ ID NO: 2.

8. The method of claim 7 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 97% sequence identity with the mature polypeptide of SEQ ID NO: 2.

9. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity comprises or consists of the amino acid sequence of SEQ ID NO: 2 or the mature polypeptide thereof; or a fragment thereof having cellulolytic enhancing activity.

10. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1.

11. The method of claim 10 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 97% sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1.

12. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or the mature polypeptide coding sequence thereof; or a subsequence thereof encoding a polypeptide fragment having cellulolytic enhancing activity.

13. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the genomic DNA 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 μg/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.

14. The method of claim 13 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the genomic DNA 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 μg/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.

15. The method of claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by the polynucleotide contained in plasmid pTr3337 which is contained in E. coli NRRL B-30878.

Assignments (1)
CONFIRMATORY LICENSE Recorded Mar 18, 2010
From: NOVOZYMES, INC.
To: ENERGY, UNITED STATES DEPARTMENT OF
Reel/Frame 024105/0747 →
Continuity (3)
Continuation 11541099 · Sep 29, 2006
Provisional Application 60722579 · Sep 30, 2005
Related Publication 20090311755A1 · Dec 17, 2009