Polypeptides having xylanase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9914946-B2
Application number	US-201715483661-A
Country	US
Kind code	B2
Filing date	Apr 10, 2017
Priority date	May 10, 2007
Publication date	Mar 13, 2018
Grant date	Mar 13, 2018

Abstract

The present invention relates to methods for degrading or converting a cellulose-containing material, comprising: treating the cellulose-containing material with an effective amount of a cellulolytic enzyme composition comprising a polypeptide having cellulolytic enhancing activity, and one or more (several) components selected from the group consisting of a CEL7 polypeptide having endoglucanase activity, a CEL12 polypeptide having endoglucanase activity, a CEL45 polypeptide having endoglucanase activity, a CEL7 polypeptide having cellobiohydrolase activity with a cellulose binding domain, and a CEL7 polypeptide having cellobiohydrolase activity without a cellulose binding domain. The present invention also relates to such cellulolytic enzyme compositions.

First claim

What is claimed is:

1. A method for producing a fermentation product, comprising:
(A) saccharifying a cellulose-containing material with a cellulolytic enzyme composition comprising an effective amount of a GH61 polypeptide having cellulolytic enhancing activity, and a CEL7 polypeptide having endoglucanase activity, a CEL12 polypeptide having endoglucanase activity, a CEL45 polypeptide having endoglucanase activity, a CEL7 polypeptide having cellobiohydrolase activity with a cellulose binding domain, a CEL7 polypeptide having cellobiohydrolase activity without a cellulose binding domain, and a polypeptide having beta-glucosidase activity;
wherein the CEL7 polypeptide having endoglucanase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) a polypeptide 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 comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) the full-length complement of (i) or (ii); and (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1;
wherein the CEL12 polypeptide having endoglucanase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 4; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 3, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 3, or (iii) the full-length complement of (i) or (ii); (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 3;
wherein the CEL45 polypeptide having endoglucanase activity is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 6; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 5, (ii) the cDNA sequence contained in the mature polypeptide coding sequence of SEQ ID NO: 5, or (iii) the full-length complement of (i) or (ii); and (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5;
wherein the CEL7 polypeptide having cellobiohydrolase activity with a cellulose binding domain is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 8; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 7, (ii) the cDNA sequence contained in the mature polypeptide coding sequence of SEQ ID NO: 7, or (iii) the full-length complement of (i) or (ii); and (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 7;
wherein the CEL7 polypeptide having cellobiohydrolase activity without a cellulose binding domain is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 10; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 9, (ii) the cDNA sequence contained in or the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 9, or (iii) the full-length complement of (i) or (ii); and (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 9;
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.; and
wherein the presence of the GH61 polypeptide having cellulolytic enhancing activity increases the degradation of a cellulose-containing material by the cellulolytic enzyme composition compared to the absence of the GH61 polypeptide having cellulolytic enhancing activity and wherein the effective amount of the GH61 polypeptide to the cellulolytic proteins of the enzyme composition is about 0.005 to about 1.0 g per g of the cellulolytic proteins;
(B) fermenting the saccharified cellulose-containing material of step (a) with one or more (several) fermenting microorganisms to produce a fermentation product; and
(C) recovering the fermentation product.

2. The method of claim 1 , wherein the CEL7 polypeptide having endoglucanase activity comprises the mature polypeptide of SEQ ID NO: 2 or a fragment thereof having endoglucanase activity, the CEL12 polypeptide having endoglucanase activity comprises the mature polypeptide of SEQ ID NO: 4 or a fragment thereof having endoglucanase activity, the CEL45 polypeptide having endoglucanase activity comprises the mature polypeptide of SEQ ID NO: 6 or a fragment thereof having endoglucanase activity, the CEL7 polypeptide having cellobiohydrolase activity with a cellulose binding domain comprises the mature polypeptide of SEQ ID NO: 8 or a fragment thereof having cellobiohydrolase activity, and the CEL7 polypeptide having cellobiohydrolase activity without a cellulose binding domain comprises the mature polypeptide of SEQ ID NO: 10 or a fragment thereof having cellobiohydrolase activity.

3. The method of claim 1 , wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having cellulolytic enhancing activity comprising [ILMV]-P-X(4,5)-G-X-Y-[ILMV]-X-R-X-[EQ]-X(4)-[HNQ] (SEQ ID NO: 31 or SEQ ID NO: 32) and [FW]-[TF]-K-[AIV] (SEQ ID NO: 41), wherein X is any amino acid, X(4,5) is any amino acid at 4 or 5 contiguous positions, and X(4) is any amino acid at 4 contiguous positions; and
(b) a GH61 polypeptide having cellulolytic enhancing activity comprising [ILMV]-P-X(4,5)-G-X-Y-[ILMV]-X-R-X-[EQ]-X(3)-A-[HNQ] (SEQ ID NO: 39 or SEQ ID NO: 40), wherein X is any amino acid, X(4,5) is any amino acid at 4 or 5 contiguous positions, and X(3) is any amino acid at 3 contiguous positions.

4. The method of claim 3 , wherein the GH61 polypeptide having cellulolytic enhancing activity comprising [ILMV]-P-X(4,5)-G-X-Y-[ILMV]-X-R-X-[EQ]-X(4)-[HNQ] (SEQ ID NO: 31 or SEQ ID NO: 32) and [FW]-[TF]-K-[AIV] (SEQ ID NO: 41) further comprises:

(SEQ ID NO: 33 or SEQ ID NO: 34)

H-X(1,2)-G-P-X(3)-[YW]-[AILMV],

(SEQ ID NO: 35)

[EQ]-X-Y-X(2)-C-X-[EHQN]-[FILV]-X-[ILV],

or

Assignees

• Novozymes Inc

Inventors

• Mcfarland Keith

Classifications

• C12Y302/01021

  Beta-glucosidase (3.2.1.21) · CPC title

• C12Y302/01004

  Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase · CPC title

• C12P7/24

  containing a carbonyl group · CPC title

• C12P7/26

  Ketones · CPC title

• C12P2203/00

  Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source · CPC title