Cellulases, the genes encoding them and uses thereof

• US 7,323,326 B2
• Filed: 02/20/2004
• Issued: 01/29/2008
• 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: Miettinen-Oinonen, Arja, Haakana, Heli, Elovainio, Minna, Joutsjoki, Vesa, Paloheimo, Marja, Lantto, Raija, Vehmaanpera, Jari, Mantyla, Arja, Suominen, Pirkko, Londesborough, John
• Primary Examiner(s): Slobodyansky; Elizabeth
• Application Number:
  US10/782,002
• Publication Number:
  US 20040142444A1
• Time in Patent Office: 1,439 Days
• Field of Search: 435/4, 435/6, 435/41, 435/69.1, 435/183, 435/200.T, 530/23.2, 530/23.7, 530/23.74, 510/114, 510/392, 510/515
• US Class Current: 435/209
• 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. An isolated and essentially homogenous polypeptide having the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

2. An isolated and essentially homogenous polypeptide having cellulase activity and an amino acid sequence which has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

3. An isolated and essentially homogenous polypeptide having cellulase activity and amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

4. An isolated and essentially homogenous polypeptide having cellulase activity and an amino acid sequence which has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

5. An enzyme extract 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 FIGS. 23A-C and SEQ ID NO;

35,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011,

(iv) a polypeptide comprising amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

(v) a polypeptide having at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35.

6. An enzyme extract preparation according to claim 5, wherein said polypeptide comprises the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

7. An enzyme extract preparation according to claim 5, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

8. An enzyme extract preparation according to claim 5, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011.

9. An enzyme extract preparation according to claim 5, wherein said polypeptide comprises amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

10. An enzyme extract preparation according to claim 5, wherein said polypeptide has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

11. An enzyme extract preparation according to claim 5, wherein said enzyme extract preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising the nucleic acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

34; and

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

12. An enzyme extract preparation according to claim 5, wherein said enzyme extract preparation is obtained by a process comprising:

(i) culturing a host cell transformed with a nucleic acid sequence encoding a polypeptide having cellulase activity and 95% identity to amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

13. An enzyme extract preparation according to claim 5, wherein said enzyme extract preparation is obtained by a process comprising:

(i) culturing a host cell transformed with a nucleic acid sequence encoding amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

14. An enzyme extract preparation according to claim 5, wherein said enzyme extract preparation is obtained by a process comprising:

(i) culturing a host cell transformed with a nucleic acid sequence encoding a polypeptide having cellulase activity and at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

15. An enzyme extract preparation according to claim 5, wherein said polypeptide is isolated and essentially homogenous.

16. An enzyme extract preparation according to claim 5, wherein said enzyme extract preparation has cellulase activity and comprises at least one cellulase of a fungal species belonging to a fungal genus selected from the group consisting of Melanocarpus and Myriococcum.

17. An enzyme extract preparation having cellulase activity according to claim 16, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces or Myriococcum sp. species represented by CBS 687.95.

18. An enzyme extract preparation having cellulase activity according to claim 17, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95 or Myriococcum sp. CBS 687.95.

19. An enzyme extract preparation according to claim 5, wherein the enzyme extract preparation is liquid and has cellulase activity.

20. An enzyme extract preparation according to claim 5, wherein the enzyme extract preparation is dry and has cellulase activity.

21. An enzyme extract preparation according to claim 5, wherein the enzyme extract preparation has cellulase activity and further comprises a surface active agent.

22. 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 FIGS. 23A-C and SEQ ID NO;

35,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011,

(iv) a polypeptide comprising amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

(v) a polypeptide having at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35.

23. A method according to claim 22, wherein said polypeptide comprises the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

24. A method according to claim 22, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

25. A method according to claim 22, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011.

26. A method according to claim 22, wherein said polypeptide comprises amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

27. A method according to claim 22, wherein said polypeptide has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

28. A method according to claim 22, 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 FIGS. 23A-C and SEQ ID NO;

34; and

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

29. A method according to claim 22, 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 FIGS. 23A-C and SEQ ID NO;

35; and

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

30. A method according to claim 22, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

31. A method according to claim 22, 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 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

32. A method according to claim 22, wherein said polypeptide is isolated and essentially homogenous.

33. A method according to claim 22, 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.

34. A method of claim 33, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces or Myriococcum sp. species represented by CBS 687.95.

35. A method of claim 34, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95 or Myriococcum sp. CBS 687.95.

36. A method according to claim 22, wherein the enzyme preparation is liquid.

37. A method according to claim 22, wherein the enzyme preparation is dry.

38. A method according to claim 22, wherein the fabric or garments is denim.

39. A method according to claim 22, wherein the enzyme preparation further comprises a surface active agent.

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

(i) a polypeptide comprising the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011,

(iv) a polypeptide comprising amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

(v) a polypeptide having at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35.

41. A method according to claim 40, wherein said polypeptide comprises the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

42. A method according to claim 40, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

43. A method according to claim 40, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011.

44. A method according to claim 40, wherein said polypeptide comprises amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

45. A method according to claim 40, wherein said polypeptide has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

46. A method according to claim 40, 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 FIGS. 23A-C and SEQ ID NO;

34; and

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

47. A method according to claim 40, 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 FIGS. 23A-C and SEQ ID NO;

35; and

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

48. A method according to claim 40, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

49. A method according to claim 40, 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 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

50. A method according to claim 40, wherein said polypeptide is isolated and essentially homogenous.

51. A method according to claim 40, 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.

52. A method of claim 51, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces or Myriococcum sp. species represented by CBS 687.95.

53. A method of claim 52, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95, or Myriococcum sp. CBS 687.95.

54. A method according to claim 40, wherein the enzyme preparation is liquid.

55. A method according to claim 40, wherein the enzyme preparation is dry.

56. A method according to claim 40, wherein the textile materials are manufactured of natural cellulose containing fibers or manmade cellulose containing fibers or are mixtures thereof.

57. A method according to claim 40, wherein the textile materials are blends of synthetic fibers and cellulose containing fibers.

58. A method according to claim 40, wherein the enzyme preparation further comprises a surface active agent.

59. 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 FIGS. 23A-C and SEQ ID NO;

35,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011,

(iv) a polypeptide comprising amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

(v) a polypeptide having at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35.

60. A method according to claim 59, wherein said polypeptide comprises the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

61. A method according to claim 59, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

62. A method according to claim 59, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011.

63. A method according to claim 59, wherein said polypeptide comprises amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

64. A method according to claim 59, wherein said polypeptide has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

65. A method according to claim 59, 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 FIGS. 23A-C and SEQ ID NO;

34; and

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

66. A method according to claim 59, 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 FIGS. 23A-C and SEQ ID NO;

35; and

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

67. A method according to claim 59, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

68. A method according to claim 59, 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 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

69. A method according to claim 59, wherein said polypeptide is isolated and essentially homogenous.

70. A method according to claim 59, 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.

71. A method of claim 70, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces or Myriococcum sp. species represented by CBS 687.95.

72. A method of claim 71, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95 or Myriococcum sp. CBS 687.95.

73. A method according to claim 59, wherein the enzyme preparation is liquid.

74. A method according to claim 59, wherein the enzyme preparation is dry.

75. A method according to claim 59, wherein the enzyme preparation further comprises a surface active agent.

76. 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 FIGS. 23A-C and SEQ ID NO;

35,

(ii) a polypeptide having at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35,

(iii) a polypeptide comprising the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011,

(iv) a polypeptide comprising amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

(v) a polypeptide having at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35.

77. A method according to claim 76, wherein said polypeptide comprises the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

78. A method according to claim 76, wherein said polypeptide has at least 95% identity to the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

79. A method according to claim 76, wherein said polypeptide comprises the amino acid sequence encoded by the DNA insert contained in DSM 11026, DSM 11013, or DSM 11011.

80. A method according to claim 76, wherein said polypeptide comprises amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

81. A method according to claim 76, wherein said polypeptide has at least 95% identity to amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO:

35.

82. A method according to claim 76, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid molecule comprising sequence set forth in FIGS. 23A-C and SEQ ID NO;

34; and

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

83. A method according to claim 76, 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 FIGS. 23A-C and SEQ ID NO;

35; and

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

84. A method according to claim 76, wherein said enzyme preparation is obtained by a process comprising:

(i) culturing a host cell transformed with the nucleic acid sequence encoding amino acids 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

85. A method according to claim 76, 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 23-452 of the amino acid sequence set forth in FIGS. 23A-C and SEQ ID NO;

35; and

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

86. A method according to claim 76, wherein said polypeptide is isolated and essentially homogenous.

87. A method according to claim 76, 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.

88. A method of claim 87, wherein the fungal species is Melanocarpus albomyces, Myriococcum albomyces or Myriococcum sp. species represented by CBS 687.95.

89. A method of claim 88, wherein the fungus is Melanocarpus albomyces, Myriococcum albomyces CBS 685.95 or Myriococcum sp. CBS 687.95.

90. A method according to claim 76, wherein the enzyme preparation is liquid.

91. A method according to claim 76, wherein the enzyme preparation is dry.

92. A method according to claim 76, wherein the enzyme preparation further comprises a surface active agent.