Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

• US 20060005279A1
• Filed: 02/04/2005
• Published: 01/05/2006
• Est. Priority Date: 02/06/2004
• Status: Active Grant

• Current Assignee: Novozymes
• Original Assignee: Novozymes Biotech, Inc. (Novozymes)
• Inventors: Ding, Hanshu, Dotson, William D., Greenier, Jennifer
• Granted Patent: US 7,271,244 B2
• Time in Patent Office: Days
• US Class Current: 800/284
• CPC Class Codes: C07K 14/37 from fungi
• CPC Class Codes: C11D 3/38645 containing cellulase
• CPC Class Codes: C12N 15/8246 Non-starch polysaccharides, e.g. cellulose, fructans, levans
• CPC Class Codes: C12N 9/2445 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• 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: C12Y 302/01091 Cellulose 1,4-beta-cellobiosidase (3.2.1.91)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

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

Claims

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

(a) a polypeptide having an amino acid sequence which has at least 70% identity with amino acids 23 to 250 of SEQ ID NO;

2;

(b) a polypeptide which is encoded by a polynucleotide which hybridizes under at least low stringency conditions with (i) nucleotides 67 to 805 of SEQ ID NO;

1, (ii) the cDNA sequence contained in nucleotides 67 to 805 of SEQ ID NO;

1, or (iii) a complementary strand of (i) or (ii); and

(c) a variant comprising a conservative substitution, deletion, and/or insertion of one or more amino acids of amino acids 23 to 250 of SEQ ID NO;

2.

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

2.

9. The polypeptide of claim 1, which consists of SEQ ID NO:

2 or a fragment thereof having cellulolytic enhancing activity.

10. The polypeptide of claim 9, which consists of SEQ ID NO:

2.

11. The polypeptide of claim 9, which consists of amino acids 23 to 250 of SEQ ID NO:

2.

16. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pDZA2-7 which is contained in E. coli NRRL B-30704.

17. An isolated polynucleotide comprising a nucleotide sequence which encodes the polypeptide of claim 1.

19. A nucleic acid construct comprising the polynucleotide of claim 17 operably linked to one or more control sequences that direct the production of the polypeptide in an expression host.

22. A method for producing the polypeptide of claim 1 comprising:

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

(b) recovering the polypeptide.

23. A method for 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.

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

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

(b) recovering the polypeptide.

39. A transgenic plant, plant part or plant cell, which has been transformed with a polynucleotide encoding the polypeptide of claim 1.

40. A detergent composition comprising the polypeptide having cellulolytic enhancing activity of claim 1, a cellulolytic activity, and a surfactant.

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

treating the cellulosic material with an effective amount of a cellulolytic protein in the presence of an effective amount 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.

53. A method for producing an organic substance, comprising:

(a) saccharifying a cellulosic material with an effective amount of a cellulolytic protein in the presence of an effective amount of the polypeptide having cellulolytic enhancing activity of claim 8, 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 fermentating microorganisms; and

(c) recovering the organic substance from the fermentation.

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21. A recombinant host cell comprising the nucleic acid construct of claim 18.

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28. An isolated polynucleotide obtained by (a) hybridizing a population of DNA under medium stringency conditions with (i) nucleotides 67 to 805 of SEQ ID NO:

1, (ii) the cDNA sequence contained in nucleotides 67 to 805 of SEQ ID NO;

1, or (iii) a complementary strand of (i) or (ii); and

(b) isolating the hybridizing polynucleotide, which encodes a polypeptide having cellulolytic enhancing activity.

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34. A nucleic acid construct comprising a gene encoding a protein operably linked to a nucleotide sequence encoding a signal peptide consisting of nucleotides 1 to 66 of SEQ ID NO:

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

36. A recombinant host cell comprising the nucleic acid construct of claim 34.

37. A method for producing a protein comprising:

(a) cultivating the recombinant host cell of claim 36 under conditions conducive for production of the protein; and

(b) recovering the protein.

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