Polypeptides having endoglucanase activity and polynucleotides encoding same

• US 8,288,615 B2
• Filed: 02/17/2012
• Issued: 10/16/2012
• Est. Priority Date: 03/20/2006
• Status: Expired due to Fees

• Current Assignee: Novozymes, Novozymes A/S (Novozymes)
• Original Assignee: Novozymes, Novozymes A/S (Novozymes)
• Inventors: Lassen, Søren Flensted, Harris, Paul, Vlasenko, Elena, Krogh, Kristian
• Primary Examiner(s): Page, Brent T
• Application Number:
  US13/399,588
• Publication Number:
  US 20120151633A1
• Time in Patent Office: 242 Days
• Field of Search: None
• US Class Current: 800/288
• 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 90% sequence identity with the mature polypeptide of SEQ ID NO;

6;

(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;

5, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO;

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

(c) a polypeptide which is encoded by a polynucleotide having at least 90% sequence identity with the mature polypeptide coding sequence of SEQ ID NO;

5.

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

6.

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

5.

4. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pCIC453 which is contained in E. coli NRRL B-30903.

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

6. The isolated polynucleotide of claim 5, comprising at least one mutation in the mature polypeptide coding sequence of SEQ ID NO:

5 in which the mutant nucleotide sequence encodes the mature polypeptide of SEQ ID NO;

6.

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

8. A recombinant host cell comprising the nucleic acid construct of claim 7.

9. 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.

10. 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.

11. 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.

12. 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.

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

14. 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.

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

treating the biomass with a composition comprising the host cell of claim 8.

16. 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.

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

6.

18. The polypeptide of claim 17, wherein the mature polypeptide is amino acids 16 to 397 of SEQ ID NO:

6.

19. The polypeptide of claim 1, which is encoded by a polynucleotide comprising or consisting of the mature polypeptide coding sequence of SEQ ID NO:

5.

20. The polypeptide of claim 19, wherein the mature polypeptide coding sequence is nucleotides 84 to 1229 of SEQ ID NO:

5.

21. The polypeptide of claim 1, comprising an amino acid sequence which has at least 95% sequence identity with the mature polypeptide of SEQ ID NO:

6.

22. The polypeptide of claim 1, comprising an amino acid sequence which has at least 97% sequence identity with the mature polypeptide of SEQ ID NO:

6.

23. The polypeptide of claim 1, comprising an amino acid sequence which has at least 98% sequence identity with the mature polypeptide of SEQ ID NO:

6.

24. The polypeptide of claim 1, comprising an amino acid sequence which has at least 99% sequence identity with the mature polypeptide of SEQ ID NO:

6.

25. The polypeptide of claim 1, which is encoded by a polynucleotide having at least 95% sequence identity with the mature polypeptide coding sequence of SEQ ID NO:

5.

26. The polypeptide of claim 1, which is encoded by a polynucleotide having at least 97% sequence identity with the mature polypeptide coding sequence of SEQ ID NO:

5.

27. The polypeptide of claim 1, which is encoded by a polynucleotide having at least 98% sequence identity with the mature polypeptide coding sequence of SEQ ID NO:

5.

28. The polypeptide of claim 1, which is encoded by a polynucleotide having at least 99% sequence identity with the mature polypeptide coding sequence of SEQ ID NO:

5.

29. The polypeptide of claim 1, which is encoded by a polynucleotide which hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO:

5, (ii) the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO;

5, or (iii) a full-length complementary strand of (i) or (ii).