Polypeptides having Cellulolytic Enhancing Activity and Polynucleotides Encoding Same

• US 20100306881A1
• Filed: 12/17/2008
• Published: 12/02/2010
• Est. Priority Date: 12/19/2007
• Status: Abandoned Application

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: Novozymes A/S (Novozymes)
• Inventors: Harris, Paul, Maiyuran, Suchindra, Brown, Kimberly
• Application Number:
  US12/745,993
• Publication Number:
  US 20100306881A1
• Time in Patent Office: Days
• US Class Current: 800/298
• CPC Class Codes: C07K 14/37 from fungi
• CPC Class Codes: C12N 9/2434 acting on beta-1,4-glucosidic bonds

Abstract

The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated 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 comprising an amino acid sequence having at least 60% identity to the mature polypeptide of SEQ ID NO;

2;

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

1, (ii) the cDNA sequence contained in the mature polypeptide coding sequence of SEQ ID NO;

1, or (iii) a full-length complementary strand of (i) or (ii);

(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 60% identity to the mature polypeptide coding sequence of SEQ ID NO;

1; and

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

2.

2. The polypeptide of claim 1, comprising or consisting of the amino acid sequence of SEQ ID NO:

2;

or a fragment thereof having cellulolytic enhancing activity.

3. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pSMai191 which is contained in E. coli NRRL B-50084.

4. An isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 1.

5. A nucleic acid construct comprising the polynucleotide of claim 4 operably linked to one or more (several) control sequences that direct the production of the polypeptide in an expression host.

6. A recombinant host cell comprising the nucleic acid construct of claim 5.

7. 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

(b) recovering the polypeptide.

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

(a) cultivating a host cell comprising a nucleic acid construct comprising a nucleotide sequence encoding the polypeptide under conditions conducive for production of the polypeptide; and

(b) recovering the polypeptide.

9. A method of producing a mutant of a parent cell, comprising disrupting or deleting a nucleotide sequence encoding the polypeptide of claim 1, which results in the mutant producing less of the polypeptide than the parent cell.

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

(a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding the polypeptide under conditions conducive for production of the polypeptide; and

(b) recovering the polypeptide.

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

12. A double-stranded inhibitory RNA (dsRNA) molecule comprising a subsequence of the polynucleotide of claim 4, wherein optionally the dsRNA is a siRNA or a miRNA molecule.

13. A method of inhibiting the expression of a polypeptide having cellulolytic enhancing activity in a cell, comprising administering to the cell or expressing in the cell a double-stranded RNA (dsRNA) molecule, wherein the dsRNA comprises a subsequence of the polynucleotide of claim 4.

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

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

18. The method of claim 17, further comprising recovering the degraded cellulosic material.

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

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

(b) fermenting the saccharified cellulosic material of step (a) with one or more fermenting microorganisms to produce the fermentation product; and

(c) recovering the fermentation product from the fermentation.

20. A method of fermenting a cellulosic material, comprising:

fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with a cellulolytic enzyme composition in the presence of a polypeptide having cellulolytic enhancing activity of claim 1 and the presence of the polypeptide having cellulolytic enhancing activity increases the degradation of the cellulosic material compared to the absence of the polypeptide having cellulolytic enhancing activity.

14. A nucleic acid construct comprising a gene encoding a protein operably linked to a nucleotide sequence encoding a signal peptide comprising or consisting of amino acids 1 to 18 of SEQ ID NO:

2, wherein the gene is foreign to the nucleotide sequence.

15. A recombinant host cell comprising the nucleic acid construct of claim 14.

16. A method of producing a protein, comprising:

(a) cultivating the recombinant host cell of claim 15 under conditions conducive for production of the protein; and

(b) recovering the protein.