Polypeptides Having Cellulolytic Enhancing Activity And Polynucleotides Encoding Same

• US 20140308705A1
• Filed: 09/20/2012
• Published: 10/16/2014
• Est. Priority Date: 09/20/2011
• Status: Abandoned Application

• Current Assignee: Novozymes
• Original Assignee: Novozymes A/S (Novozymes)
• Inventors: Morant, Marc, Shaghasi, Tarana
• Application Number:
  US14/345,886
• Publication Number:
  US 20140308705A1
• Time in Patent Office: Days
• US Class Current: 435/69.8
• CPC Class Codes: C07K 14/37 from fungi
• CPC Class Codes: C12N 15/8246 Non-starch polysaccharides, e.g. cellulose, fructans, levans
• 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: C12N 9/2445 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C12P 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• 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 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source ethanol C12P7/10
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The present invention relates to isolated polypeptides having cellulolytic enhancing activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Claims

1. An isolated polypeptide having cellulolytic enhancing activity, selected from the group consisting of:

(a) a polypeptide having at least 75% sequence identity to the mature polypeptide of SEQ ID NO;

2;

at least 90% to the mature polypeptide of SEQ ID NO;

4;

or at least 85% sequence identity to the mature polypeptide of SEQ ID NO;

6;

(b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO;

1, SEQ ID NO;

3, or SEQ ID NO;

5, or (ii) the full-length complement of (i);

(c) a polypeptide encoded by a polynucleotide having at least 75% sequence identity to the mature polypeptide coding sequence of SEQ ID NO;

1;

at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO;

3;

or at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO;

5;

(d) a variant of the mature polypeptide of SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6 comprising a substitution, deletion, and/or insertion at one or more positions;

(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has cellulolytic enhancing activity; and

(f) a polypeptide comprising or consisting of SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6 or the mature polypeptide of SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6.

4. A nucleic acid construct comprising an isolated polynucleotide encoding the polypeptide of claim 1, wherein the polynucleotide is operably linked to control sequences that direct expression of the polynucleotide in a suitable host cell and one or more of the control sequences are foreign to the polynucleotide.

5. A method of producing the polypeptide of claim 1, comprising:

(a) cultivating a cell, which in its wild-type form produces the polypeptide, under conditions conducive for production of the polypeptide; and

optionally

(b) recovering the polypeptide.

6. A method of producing a polypeptide having cellulolytic enhancing activity, comprising:

(a) cultivating a recombinant host cell comprising the polynucleotide of claim 4 under conditions conducive for production of the polypeptide; and

optionally

(b) recovering the polypeptide.

7. A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding the polypeptide of claim 1.

8. A method of producing a polypeptide having cellulolytic enhancing activity, comprising:

(a) cultivating the transgenic plant or plant cell of claim 7 under conditions conducive for production of the polypeptide; and

optionally

(b) recovering the polypeptide.

9. A method of producing a mutant of a parent cell, comprising inactivating a polynucleotide encoding the polypeptide of claim 1, which results in the mutant producing less of the polypeptide than the parent cell.

12. A process for degrading a cellulosic material, comprising:

treating the cellulosic material with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of claim 1.

13. A process for producing a fermentation product, comprising:

(a) saccharifying a cellulosic material with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of claim 1;

(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.

14. A process of fermenting a cellulosic material, comprising:

fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of claim 1.

15. The process of claim 14, wherein the fermenting of the cellulosic material produces a fermentation product.

16. A whole broth formulation or cell culture composition comprising the polypeptide of claim 1.

2. An isolated polypeptide comprising a catalytic domain selected from the group consisting of:

(a) a catalytic domain having at least 90% sequence identity to amino acids 20 to 252 of SEQ ID NO;

4;

(b) a catalytic domain encoded by a polynucleotide that hybridizes under at least high stringency conditions with nucleotides 58 to 756 of SEQ ID NO;

3 or the full-length complement thereof;

(c) a catalytic domain encoded by a polynucleotide having at least 90% sequence identity to nucleotides 58 to 756 of SEQ ID NO;

3;

(d) a variant of amino acids 20 to 252 of SEQ ID NO;

4 comprising a substitution, deletion, and/or insertion at one or more positions; and

(e) a fragment of the catalytic domain of (a), (b), (c), or (d) that has cellulolytic enhancing activity.

3. An isolated polypeptide comprising a cellulose binding domain operably linked to a catalytic domain, wherein the binding domain is selected from the group consisting of:

(a) a cellulose binding domain having at least 90% sequence identity to amino acids 309 to 344 of SEQ ID NO;

4;

(b) a cellulose binding domain encoded by a polynucleotide that hybridizes under at least very high stringency conditions with nucleotides 925 to 1032 of SEQ ID NO;

3 or the full-length complement thereof;

(c) a cellulose binding domain encoded by a polynucleotide having at least 90% sequence identity to nucleotides 925 to 1032 of SEQ ID NO;

3;

(d) a variant of amino acids 309 to 344 of SEQ ID NO;

4 comprising a substitution, deletion, and/or insertion at one or more positions; and

(e) a fragment of the cellulose binding domain of (a), (b), (c), or (d) that has binding activity.

10. An isolated polynucleotide encoding a signal peptide comprising or consisting of amino acids 1 to 19 of SEQ ID NO:

2, amino acids 1 to 19 of SEQ ID NO;

4, or amino acids 1 to 21 of SEQ ID NO;

6.

11. A method of producing a protein, comprising:

(a) cultivating a recombinant host cell comprising a gene encoding a protein operably linked to the polynucleotide of claim 10, wherein the gene is foreign to the polynucleotide encoding the signal peptide, under conditions conducive for production of the protein; and

optionally

(b) recovering the protein.