Polypeptides having endoglucanase activity and polynucleotides encoding same

• US 20110047652A1
• Filed: 03/09/2007
• Published: 02/24/2011
• Est. Priority Date: 03/20/2006
• Status: Active Grant

• Current Assignee: Novozymes, Novozymes A/S (Novozymes)
• Original Assignee: Novozymes, Novozymes A/S (Novozymes)
• Inventors: Lassen, Soren Flensted, Vlasenko, Elena, Harris, Paul, Krogh, Kristian Bertel Romer Morkeberg
• Granted Patent: US 8,143,482 B2
• Time in Patent Office: Days
• US Class Current: 800/298
• CPC Class Codes: C12N 9/2434 acting on beta-1,4-glucosidic bonds
• 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: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

Abstract

The present invention relates to isolated polypeptides having endoglucanase 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 for producing and using the polypeptides.

Claims

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

(a) a polypeptide comprising an amino acid sequence which has at least 80% identity with the mature polypeptide of SEQ ID NO;

4 or SEQ ID NO;

10, at least 85% identity with the mature polypeptide of SEQ ID NO;

6, or at least 75% identity with the mature polypeptide of SEQ ID NO;

8;

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

3, SEQ ID NO;

5,SEQ ID NO;

7, or SEQ ID NO;

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

3 or SEQ ID NO;

9 or the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO;

5 or SEQ ID NO;

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

(c) a polypeptide which is encoded by a polynucleotide having at least 80% identity with the mature polypeptide coding sequence of SEQ ID NO;

3 or SEQ ID NO;

9, at least 85% identity with the mature polypeptide coding sequence of SEQ ID NO;

5, or at least 75% identity with the mature polypeptide coding sequence of SEQ ID NO;

7.

3. The polypeptide of claim 1, which comprises or consists of the amino acid sequence of SEQ ID NO:

4, SEQ ID NO;

6, SEQ ID NO;

8, or SEQ ID NO;

10;

or a fragment thereof having endoglucanase activity.

8. The polypeptide of claim 1, which is encoded by a polynucleotide comprising or consisting of SEQ ID NO:

3, SEQ ID NO;

5, SEQ ID NO;

7, or SEQ ID NO;

9;

or a subsequence thereof which encodes a polypeptide fragment having endoglucanase activity.

12. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pCIC161 which is contained in E. coli NRRL B-30902, pCIC453 which is contained in E. coli NRRL B-30903, pCIC486 which is contained in E. coli NRRL B-30904, or pPBCel5C which is contained in E. coli NRRL B-30900N.

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

18. A nucleic acid construct comprising the polynucleotide of claim 16 operably linked to one or more control sequences that direct the production of the polypeptide in an expression host.

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

21. 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.

22. 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.

23. A method for producing a mutant of a parent cell, which comprises 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.

37. 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 endoglucanase activity under conditions conducive for production of the polypeptide; and

(b) recovering the polypeptide.

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

39. A method of degrading or converting a cellulosic material, comprising:

treating the cellulosic material with a composition comprising an effective amount of a polypeptide having endoglucanase activity of claim 1.

44. A method of degrading or converting a cellulosic material, comprising:

treating the biomass with a composition comprising the host cell of claim 20.

48. A method of producing a substance, comprising:

(a) saccharifying a cellulosic material with a composition comprising an effective amount of a polypeptide having endoglucanase activity of claim 1, (b) fermenting the saccharified cellulosic material of step (a) with one or more fermentating microorganisms; and

(c) recovering the substance from the fermentation.

2. (canceled)

4. (canceled)

5. (canceled)

6. (canceled)

7. (canceled)

9. (canceled)

10. (canceled)

11. (canceled)

13. (canceled)

14. (canceled)

15. (canceled)

17. (canceled)

19. (canceled)

24. (canceled)

25. (canceled)

26. (canceled)

27. (canceled)

28. (canceled)

29. (canceled)

30. (canceled)

31. (canceled)

32. (canceled)

33. A nucleic acid construct comprising a gene encoding a protein operably linked to one or both of a first nucleotide sequence encoding a signal peptide comprising or consisting of amino acids 1 to 17 of SEQ ID NO:

4, amino acids 1 to 15 of SEQ ID NO;

6, amino acids 1 to 21 of SEQ ID NO;

8, or amino acids 1 to 16 of SEQ ID NO;

10, and a second nucleotide sequence encoding a propeptide comprising or consisting of amino acids 17 to 24 of SEQ ID NO;

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

35. A recombinant host cell comprising the nucleic acid construct of claim 33.

36. A method for producing a protein comprising:

(a) cultivating the recombinant host cell of claim 35 under conditions conducive for production of the protein; and

(b) recovering the protein.

34. (canceled)

40. (canceled)

41. (canceled)

42. (canceled)

43. (canceled)

45. (canceled)

46. (canceled)

47. (canceled)

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51. (canceled)

52. (canceled)

53. A method of inhibiting expression of a polypeptide 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 or portion of a polynucleotide encoding the mature polypeptide of SEQ ID NO:

4, SEQ ID NO;

6, SEQ ID NO;

8, or SEQ ID NO;

10.

54. (canceled)

55. A double-stranded RNA (dsRNA) molecule for inhibiting expression of a polypeptide in a cell, wherein the dsRNA comprises a subsequence or portion of a polynucleotide encoding the mature polypeptide of SEQ ID NO:

4, SEQ ID NO;

6, SEQ ID NO;

8, or SEQ ID NO;

10.