Cellulases, the genes encoding them and uses thereof

• US 7,273,748 B2
• Filed: 04/16/2004
• Issued: 09/25/2007
• Est. Priority Date: 10/17/1995
• Status: Expired due to Fees

• Current Assignee: AB Enzymes Oy (Garfield Weston Foundation)
• Original Assignee: AB Enzymes Oy (Garfield Weston Foundation)
• Inventors: Paloheimo, Marja, Lantto, Raija, Vehmaanpera, Jari, Mantyla, Arja, Suominen, Pirkko, Miettinen-Oinonen, Arja, Haakana, Heli, Elovainio, Minna, Joutsjoki, Vesa, Londesborough, John
• Primary Examiner(s): Prouty; Rebecca E.
• Assistant Examiner(s): RAGHU, GANAPATHIRAM
• Application Number:
  US10/825,378
• Publication Number:
  US 20040185498A1
• Time in Patent Office: 1,257 Days
• Field of Search: None
• US Class Current: 435/263
• CPC Class Codes: C11D 3/38645 containing cellulase
• 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

Genes encoding novel cellulases, and a gene encoding a protein that facilitates the action of such novel cellulases, the novel cellulases and a protein that facilitates the action of such cellulases, and enzyme preparations containing such proteins are described. The native hosts and the culture medium of said hosts containing said novel cellulases are also disclosed. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

Claims

1. A method for biostoning comprising adding an enzyme preparation comprising a polypeptide having cellulase activity to cotton containing fabric or garments, wherein said polypeptide is selected from the group consisting of:

(i) a polypeptide comprising the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012,

(iv) a polypeptide comprising amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31; and

(v) a polypeptide having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31.

2. A method according to claim 1, wherein said polypeptide comprises the amino acid sequence set forth in FIG. 19 and SEQ II) NO:

31.

3. A method according to claim 1, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

4. A method according to claim 1, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012.

5. A method according to claim 1, wherein said polypeptide comprises amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

6. A method according to claim 1, wherein said polypeptide has at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

7. A method according to claim 1, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising the sequence set forth in FIG. 19 (SEQ ID NO;

     30); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

8. A method according to claim 1, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

9. A method according to claim 1, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

10. A method according to claim 1, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

11. A method according to claim 1, wherein said polypeptide is isolated and essentially homogenous.

12. A method according to claim 1, wherein said enzyme preparation comprises at least one cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

13. A method of claim 12, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces, or Myriococcum sp. species represented by CBS 687.95.

14. A method of claim 13, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. CBS 687.95.

15. A method according to claim 1, wherein the enzyme preparation is liquid.

16. A method according to claim 1, wherein the enzyme preparation is dry.

17. A method according to claim 1, wherein the fabric or garments is denim.

18. A method according to claim 1, wherein the enzyme preparation further comprises a surface active agent.

72. A method according to claims 1, 19, 38, or 55, wherein said enzyme preparation further comprises at least one other cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

73. A method according to claims 1, 19, 38, or 55, wherein said enzyme preparation is a partially or completely purified Melanocarpus cellulase fraction.

74. A method according to claims 1, 19, 38, or 55, wherein said enzyme preparation is a culture supernatant comprising the cellulases derived from Melanocarpus albomyces.

19. A method for biofinishing comprising adding an enzyme preparation comprising a polypeptide having cellulase activity to textile materials like fabrics or garments or yarn, wherein said polypeptide is selected from the group consisting of:

(i) a polypeptide comprising the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012,

(iv) a polypeptide comprising amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31; and

(v) a polypeptide having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31.

20. A method according to claim 19, wherein said polypeptide comprises the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

21. A method according to claim 19, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

22. A method according to claim 19, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012.

23. A method according to claim 19, wherein said polypeptide comprises amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

24. A method according to claim 19, wherein said polypeptide has at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

25. A method according to claim 19, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising the sequence set forth in FIG. 19 (SEQ ID NO;

     30); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

26. A method according to claim 19, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

27. A method according to claim 19, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

28. A method according to claim 19, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID) NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

29. A method according to claim 19, wherein said polypeptide is isolated and essentially homogenous.

30. A method according to claim 19, wherein said enzyme preparation comprises at least one cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

31. A method of claim 30, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces, or Myriococcum sp. species represented by CBS 687.95.

32. A method of claim 31, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. CBS 687.95.

33. A method according to claim 19, wherein the enzyme preparation is liquid.

34. A method according to claim 19, wherein the enzyme preparation is dry.

35. A method according to claim 19, wherein the textile materials are manufactured of natural cellulose containing fibers or manmade cellulose containing fibers or are mixtures thereof.

36. A method according to claim 19, wherein the textile materials are blends of synthetic fibers and cellulose containing fibers.

37. A method according to claim 19, wherein the enzyme preparation further comprises a surface active agent.

38. A method for treating wood-derived pulp or fiber, comprising adding an enzyme preparation comprising a polypeptide having cellulase activity to wood-derived mechanical or chemical pulp or secondary fiber, wherein said polypeptide is selected from the group consisting of:

(i) a polypeptide comprising the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012,

(iv) a polypeptide comprising amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31; and

(v) a polypeptide having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31.

39. A method according to claim 38, wherein said polypeptide comprises the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

40. A method according to claim 38, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

41. A method according to claim 38, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012.

42. A method according to claim 38, wherein said polypeptide comprises amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

43. A method according to claim 38, wherein said polypeptide has at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

44. A method according to claim 38, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising the sequence set forth in FIG. 19 (SEQ ID NO;

     30); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

45. A method according to claim 38, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

46. A method according to claim 38, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

47. A method according to claim 38, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

48. A method according to claim 38, wherein said polypeptide is isolated and essentially homogenous.

49. A method according to claim 38, wherein said enzyme preparation comprises at least one cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

50. A method of claim 49, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces, or Myriococcum sp. species represented by CBS 687.95.

51. A method of claim 50, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. CBS 687.95.

52. A method according to claim 38, wherein the enzyme preparation is liquid.

53. A method according to claim 38, wherein the enzyme preparation is dry.

54. A method according to claim 38, wherein the enzyme preparation further comprises a surface active agent.

55. A method for improving the quality of animal feed, comprising treating plant material with an enzyme preparation comprising a polypeptide having cellulase activity, wherein said polypeptide is selected from the group consisting of:

(i) a polypeptide comprising the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012,

(iv) a polypeptide comprising amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31; and

(v) a polypeptide having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO;

31.

56. A method according to claim 55, wherein said polypeptide comprises the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

57. A method according to claim 55, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

58. A method according to claim 55, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in the vector transformed into DSM 11024 or DSM 11012.

59. A method according to claim 55, wherein said polypeptide comprises amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

60. A method according to claim 55, wherein said polypeptide has at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 and SEQ ID NO:

31.

61. A method according to claim 55, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising the sequence set forth in FIG. 19 (SEQ ID NO;

     30); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

62. A method according to claim 55, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

63. A method according to claim 55, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

64. A method according to claim 55, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding a cellulase having at least 95% identity to amino acids 22-235 of the amino acid sequence set forth in FIG. 19 (SEQ ID NO;

     31); and

(ii) separating said host cell from the culture medium and obtaining the supernatant having cellulase activity.

65. A method according to claim 55, wherein said polypeptide is isolated and essentially homogenous.

66. A method according to claim 55, wherein said enzyme preparation comprises at least one cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

67. A method of claim 66, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. represented by CBS 687.95.

68. A method of claim 67, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. CBS 687.95.

69. A method according to claim 55, wherein the enzyme preparation is liquid.

70. A method according to claim 55, wherein the enzyme preparation is dry.

71. A method according to claim 55, wherein the enzyme preparation further comprises a surface active agent.