Polypeptides having cellobiohydrolase activity and polynucleotides encoding same

• US 20060218671A1
• Filed: 01/06/2006
• Published: 09/28/2006
• Est. Priority Date: 01/06/2005
• Status: Active Grant

• Current Assignee: Novozymes
• Original Assignee: Novozymes
• Inventors: Merino, Sandra, Harris, Paul, Brown, Kimberly, Lopez De Leon, Alfredo
• Granted Patent: US 7,220,565 B2
• Time in Patent Office: Days
• US Class Current: 800/284
• CPC Class Codes: C11D 3/38636 containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase
• 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 302/01091 Cellulose 1,4-beta-cellobiosidase (3.2.1.91)

Abstract

The present invention relates to isolated polypeptides having cellobiohydrolase 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 cellobiohydrolase 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;

2;

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

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

2.

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

2;

or a fragment thereof having cellobiohydrolase activity.

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

2;

or a fragment thereof having cellobiohydrolase activity.

16. The polypeptide of claim 1, which is encoded by the polynucleotide contained in plasmid pTter6A which is contained in E. coli NRRL B-30802.

17. The polypeptide of claim 1, wherein the mature polypeptide is amino acids 18 to 481 of SEQ ID NO:

2.

18. The polypeptide of claim 1, wherein the mature polypeptide coding sequence is nucleotides 52 to 1443 of SEQ ID NO:

1.

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

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

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

24. 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.

25. 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.

26. 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.

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

42. A detergent composition comprising a polypeptide having cellobiohydrolase activity of claim 1 and a surfactant.

43. A method for degrading or converting cellulose- and hemicellulose-containing biomass, comprising treating the biomass with an effective amount of a polypeptide having cellobiohydrolase activity of claim 1 and recovering the degraded biomass.

45. A method for degrading or converting a cellulose- and hemicellulose-containing biomass, comprising treating the biomass with a host cell of claim 23 and recovering the degraded biomass.

2. (canceled)

3. (canceled)

4. (canceled)

5. (canceled)

6. (canceled)

8. (canceled)

9. (canceled)

11. (canceled)

12. (canceled)

13. (canceled)

14. (canceled)

15. (canceled)

20. (canceled)

22. (canceled)

27. (canceled)

28. (canceled)

29. (canceled)

30. An isolated polynucleotide obtained by (a) hybridizing a population of DNA under medium-high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO:

1, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence 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 cellobiohydrolase activity.

31. (canceled)

32. (canceled)

33. (canceled)

34. (canceled)

35. (canceled)

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

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

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

39. A method for producing a protein comprising:

(a) cultivating the recombinant host cell of claim 38 under conditions conducive for production of the protein; and

(b) recovering the protein.

37. (canceled)

40. (canceled)

44. (canceled)

46. (canceled)