Polypeptides having endoglucanase activity and polynucleotides encoding same

• US 8,063,267 B2
• Filed: 03/30/2007
• Issued: 11/22/2011
• Est. Priority Date: 03/30/2006
• Status: Expired due to Fees

• Current Assignee: Novozymes, Novozymes A/S (Novozymes)
• Original Assignee: Novozymes, Novozymes A/S (Novozymes)
• Inventors: Harris, Paul, Vlasenko, Elena, Kauppinnen, Marcus Sakari, Zaretsky, Elizabeth, Teter, Sarah, Brown, Kimberly
• Primary Examiner(s): Page, Brent T
• Application Number:
  US12/294,573
• Publication Number:
  US 20090328259A1
• Time in Patent Office: 1,698 Days
• Field of Search: None
• US Class Current: 800/288
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/5431 IL-11
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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 9/244 Endo-1,3(4)-beta-glucanase (3.2.1.6)
• 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 21/00 Preparation of peptides or proteins single cell protein C12N1/00
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

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 having at least 95% sequence identity with the mature polypeptide of SEQ ID NO;

2;

(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;

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

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

(c) a polypeptide which is encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity with the mature polypeptide coding sequence of SEQ ID NO;

1.

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

2.

3. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pPH50 which is contained in E. coli NRRL B-30899.

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

5. A nucleic acid construct comprising the polynucleotide of claim 4 operably linked to one or more 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 for 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 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.

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

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

(b) recovering the polypeptide.

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

11. 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.

12. The method of claim 11, wherein the composition further comprises an effective amount of an endo-1,4-beta-glucanase, exo-1,4-beta-D-glucanase, and/or beta-D-glucosidase.

13. The method of claim 11, further comprising recovering the degraded or converted cellulosic material.

14. 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.

15. The method of claim 14, wherein the substance is an alcohol, organic acid, ketone, amino acid, or gas.

16. The polypeptide of claim 1, comprising an amino acid sequence having at least 97% sequence identity to the mature polypeptide of SEQ ID NO:

2.

17. The polypeptide of claim 1, comprising an amino acid sequence having at least 98% sequence identity to the mature polypeptide of SEQ ID NO:

2.

18. The polypeptide of claim 1, comprising an amino acid sequence having at least 99% sequence identity to the mature polypeptide of SEQ ID NO:

2.

19. The polypeptide of claim 1, wherein the mature polypeptide is amino acids 23 to 464 of SEQ ID NO:

2.

20. The polypeptide of claim 1, which is encoded by a polynucleotide that hybridizes under at least very high 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).

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

1.

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

1.

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

1.

24. The polypeptide of claim 1, wherein the mature polypeptide coding sequence is nucleotides 79 to 1461 of SEQ ID NO:

1.

25. A composition comprising the polypeptide of claim 1.