Methods for enhancing the degradation or conversion of cellulosic material

• US 20070077630A1
• Filed: 09/29/2006
• Published: 04/05/2007
• Est. Priority Date: 09/30/2005
• Status: Active Grant

• Current Assignee: Novozymes
• Original Assignee: Novozymes
• Inventors: Harris, Paul, Ding, Hanshu, Rey, Michael
• Granted Patent: US 7,608,689 B2
• Time in Patent Office: Days
• US Class Current: 435/105
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12P 13/04 Alpha- or beta- amino acids other amino acids C12P13/005
• CPC Class Codes: C12P 19/14 produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase , e.g. by cellulase, hemicellulase
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12P 7/26 Ketones
• CPC Class Codes: C12P 7/40 containing a carboxyl group including Peroxycarboxylic acids
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: C12Y 302/01021 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C13K 1/02 obtained by saccharification of cellulosic materials manufacture of fodder A23K10/32
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

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

Claims

1. A method for degrading or converting a cellulosic material, comprising:

treating the cellulosic material with an effective amount of one or more cellulolytic proteins in the presence of an effective amount of a 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;

(a) a polypeptide comprising an amino acid sequence which has at least 70% identity with the mature polypeptide of SEQ ID NO;

2;

(b) a polypeptide encoded by a polynucleotide which hybridizes under at least medium 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) a complementary strand of (i) or (ii);

(c) a polypeptide comprising [ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(3)-A-[HNQ], 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; and

(d) a variant comprising a conservative substitution, deletion, and/or insertion of one or more amino acids of the mature polypeptide of SEQ ID NO;

2.

5. 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, esterase, protease, laccase, peroxidase, or a mixture thereof.

9. The method of claim 1, further comprising recovering the degraded cellulosic material.

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

20. 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.

25. 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.

26. The method of claim 1, wherein the mature polypeptide coding sequence of SEQ ID NO:

1 is nucleotides 77 to 766 of SEQ ID NO;

1.

27. The method of claims 1, wherein the mature polypeptide of SEQ ID NO:

2 is amino acids 20 to 249 of SEQ ID NO;

2.

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29. A method for producing an substance, 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 which has at least 70% identity with the mature polypeptide of SEQ ID NO;

2;

(ii) a polypeptide encoded by a polynucleotide which hybridizes under at least medium stringency conditions with (a) the mature polypeptide coding sequence of SEQ ID NO;

1, (b) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO;

1, or (c) a complementary strand of (a) or (b);

(iii) a polypeptide comprising [ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(3)-A-[HNQ], 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; and

(iv) a variant comprising a conservative substitution, deletion, and/or insertion of one or more amino acids of the mature polypeptide of SEQ ID NO;

2;

(B) fermenting the saccharified cellulosic material of step (a) with one or more fermentating microorganisms; and

(C) recovering the substance from the fermentation.

33. The method of claim 29, further comprising treating the cellulosic material with an effective amount of one or more enzymes selected from the group consisting of a hemicellulase, esterase, protease, laccase, peroxidase, or a mixture thereof.

36. The method of claim 29, wherein the substance is an alcohol, organic acid, ketone, amino acid, or gas.

42. The method of claim 29, wherein the cellulolytic protein and/or the polypeptide having cellulolytic enhancing activity are in the form of a fermentation broth with or without cells.

50. The method of claim 29, 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.

55. The method of claim 29, 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.

56. The method of claim 29, wherein the mature polypeptide coding sequence of SEQ ID NO:

1 is nucleotides 77 to 766 of SEQ ID NO;

1.

57. The method of claims 29, wherein the mature polypeptide of SEQ ID NO:

2 is amino acids 20 to 249 of SEQ ID NO;

2.

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59. A detergent composition comprising a polypeptide having cellulolytic enhancing activity is selected from the group consisting of:

(a) a polypeptide comprising an amino acid sequence which has at least 70% identity with the mature polypeptide of SEQ ID NO;

2;

(b) a polypeptide encoded by a polynucleotide which hybridizes under at least medium 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) a complementary strand of (i) or (ii);

(c) a polypeptide comprising [ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(3)-A-[HNQ], 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; and

(d) a variant comprising a conservative substitution, deletion, and/or insertion of one or more amino acids of the mature polypeptide of SEQ ID NO;

2.

67. The detergent composition of claim 59, 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.

72. The detergent composition of claim 59, 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.

73. The detergent composition of claim 59, wherein the mature polypeptide coding sequence of SEQ ID NO:

1 is nucleotides 77 to 766 of SEQ ID NO;

1.

74. The detergent composition of claim 59, wherein the mature polypeptide of SEQ ID NO:

2 is amino acids 20 to 249 of SEQ ID NO;

2.

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