Polypeptides having endoglucanase activity and polynucleotides encoding same

• US 9,428,742 B2
• Filed: 08/05/2013
• Issued: 08/30/2016
• 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, Soren Flensted, Harris, Paul, Vlasenko, Elena, Krogh, Kristian
• Primary Examiner(s): Page, Brent T
• Application Number:
  US13/959,577
• Publication Number:
  US 20130340122A1
• Time in Patent Office: 1,121 Days
• Field of Search: None
• US Class Current: 1/1
• 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. A nucleic acid construct comprising a polynucleotide encoding a polypeptide having endoglucanase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide in an expression host, and wherein the polypeptide is selected from:

(a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO;

10;

(b) a polypeptide which is encoded by a polynucleotide which hybridizes under at least high stringency conditions with (i) nucleotides 73 to 1468 of SEQ ID NO;

9, (ii) the cDNA sequence contained in nucleotides 73 to 1468 of SEQ ID NO;

9, 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 nucleotides 73 to 1468 of SEQ ID NO;

9.

2. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of SEQ ID NO:

10.

3. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity is encoded by a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO:

9.

4. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity is encoded by the polynucleotide contained in plasmid pPBCel5C which is contained in E. coli NRRL B-30900N.

5. A recombinant expression vector comprising the nucleic acid construct of claim 1.

6. A recombinant host cell comprising the nucleic acid construct of claim 1.

10. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of amino acids 25 to 421 of SEQ ID NO:

10.

11. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

12. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 95% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

13. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 97% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

14. The nucleic acid construct of claim 1, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 99% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

7. A method for producing a polypeptide having endoglucanase activity, said method comprising:

(i) cultivating a recombinant host cell comprising a polynucleotide encoding a polypeptide having endoglucanase activity under conditions conducive for production of the polypeptide; and

(ii) recovering the polypeptide having endoglucanase activity;

wherein the polypeptide having endoglucanase activity is selected from;

(a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO;

10;

(b) a polypeptide which is encoded by a polynucleotide which hybridizes under at least high stringency conditions with (i) nucleotides 73 to 1468 of SEQ ID NO;

9, (ii) the cDNA sequence contained in nucleotides 73 to 1468 of SEQ ID NO;

9, 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 nucleotides 73 to 1468 of SEQ ID NO;

9.

15. The method of claim 7, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

16. The method of claim 7, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 95% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

17. The method of claim 7, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of amino acids 25 to 421 of SEQ ID NO:

10.

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

(i) treating the cellulosic material with a composition comprising a polypeptide having endoglucanase activity; and

(ii) recovering the degraded or converted cellulosic material;

wherein the polypeptide having endoglucanase activity is selected from;

(a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO;

10;

(b) a polypeptide which is encoded by a polynucleotide which hybridizes under at least high stringency conditions with (i) nucleotides 73 to 1468 of SEQ ID NO;

9, (ii) the cDNA sequence contained in nucleotides 73 to 1468 of SEQ ID NO;

9, 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 nucleotides 73 to 1468 of SEQ ID NO;

9.

18. The method of claim 8, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

19. The method of claim 8, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 95% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

20. The method of claim 8, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of amino acids 25 to 421 of SEQ ID NO:

10.

9. A method of producing a substance, said method comprising:

(i) saccharifying a cellulosic material with a composition, wherein the composition comprises a polypeptide having endoglucanase activity;

(ii) fermenting the saccharified cellulosic material of step (i) with one or more fermenting microorganisms; and

(iii) recovering the substance from the fermentation;

wherein the polypeptide having endoglucanase activity is selected from;

(a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO;

10;

(b) a polypeptide which is encoded by a polynucleotide which hybridizes under at least high stringency conditions with (i) nucleotides 73 to 1468 of SEQ ID NO;

9, (ii) the cDNA sequence contained in nucleotides 73 to 1468 of SEQ ID NO;

9, 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 nucleotides 73 to 1468 of SEQ ID NO;

9.

21. The method of claim 9, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

22. The method of claim 9, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 95% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

23. The method of claim 9, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of amino acids 25 to 421 of SEQ ID NO:

10.

24. A recombinant host cell transformed with a nucleic acid construct or expression vector comprising a polynucleotide encoding a polypeptide having endoglucanase activity, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 90% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

25. The recombinant host cell of claim 24, wherein the polypeptide having endoglucanase activity comprises an amino acid sequence which has at least 95% sequence identity with amino acids 25 to 421 of SEQ ID NO:

10.

26. The recombinant host cell of claim 24, wherein the polypeptide having endoglucanase activity comprises or consists of the amino acid sequence of amino acids 25 to 421 of SEQ ID NO:

10.