Cellulase compositions and methods of use

• US 5,811,381 A
• Filed: 10/10/1996
• Issued: 09/22/1998
• Est. Priority Date: 10/10/1996
• Status: Expired due to Term

• Current Assignee: Danisco USA Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Mark A. Emalfarb
• Inventors: Solovjeva, Irina Vladimirovna, Olson, Philip T., Okounev, Oleg Nicolaevich, Sinitsyn, Arkady Panteleimonovich, Burlingame, Richard P., Ben-Bassat, Arie, Chernoglazov, Vladimir Mikhaylovich, Emalfarb, Mark Aaron
• Primary Examiner(s): Lau, Kawai
• Application Number:
  US08/731,170
• Time in Patent Office: 712 Days
• Field of Search: 435/200, 435/209, 510/320, 510/392, 424/94.61
• US Class Current: 510/320
• CPC Class Codes: C11D 2111/12 Soft surfaces, e.g. textile
• CPC Class Codes: C11D 3/38645 containing cellulase
• CPC Class Codes: C12N 1/145 Fungal isolates
• 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: C12R 2001/645 Fungi ; Processes using fungi
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: D06L 4/40 using enzymes
• CPC Class Codes: D06M 16/003 with enzymes or microorganisms
• CPC Class Codes: D06M 2101/06 cellulosic
• CPC Class Codes: D06P 5/02 After-treatment
• CPC Class Codes: D06P 5/158 with other compounds

Abstract

The subject invention relates to novel compositions of neutral and/or alkaline cellulase and methods for obtaining neutral and/or alkaline cellulase compositions from Chrysosporium cultures, in particular Chrysosporium lucknowense. This invention also provides mutants and methods of generating mutants of Chrysosporium capable of producing neutral and/or alkaline cellulose. This invention also relates to the genes encoding the enzymes comprising the neutral and/or alkaline cellulase composition. In addition, this invention provides methods of culturing Chrysosporium to produce neutral and/or alkaline cellulases. The neutral and/or alkaline cellulase compositions of the subject invention can be used in a variety of processes including stone washing of clothing, detergent processes, deinking and biobleaching of paper & pulp and treatment of waste streams.

Claims

1. A neutral and/or alkaline cellulase comprise obtained by a method which comprises:

(a) growing a wild type Chrysosporium lucknowense culture on a suitable medium; and

(b) isolating said neutral and/or alkaline cellulase composition from said Chrysosporium lucknowense culture.

2. The composition of claim 1, wherein said Chrysosporium lucknowense is Chrysosporium lucknowense Garg 27K, accession number VKM F-3500D.

3. A composition for the enzymatic treatment of cellulosic fibers or fabrics, comprising a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild type fungus of the genus Chrysoporium, said compostion having a pH between about 8.0 and about 12.0.

4. The composition of claim 3, wherein the fungus is selected from the group consisting of Chyrosoporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysoporium keratinophilum, Chrysosporium lobatum, Chrusoporium merdarium, Chrusoporium queenslandicum, and Chrysosporium tropictum.

5. The composition of claim 4, wherein the fungus is Chrysosporium lucknowense.

6. The composition of claim 3, wherein the cellulase is isolated or obtained form a wild-type fungus of the genus Chrysosporium.

7. The composition of claim 4, wherein the cellulase is isolated or obtained from a wild-type fungus selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenlandicum and Chrysosporium tropicum.

8. The composition of claim 5, wherein the cellulase is isolated or obtained from a wild-type fungus Chrysosporium lucknowense.

9. A composition as described in any one of claims 3-8 further comprising one or more components that enhance the performance of the cellulase, wherein the components are selected from the group consisting of pumice stones, abrasives, softeners, solvents, preservatives, bleaching agents, bluing agents, flourescent dyes, antioxidants, solubilizers, detergents, surfactants, enzymes, builders, anti-redoposition agents, buffers, caking inhibitors, masking agents for factors inhibiting the cellulase activity, and cellulase activators.

10. A composition as described in any one of claims 3-8, wherein the pH is between 10.0 and about 11.0.

11. The composition of claim 9, wherein the pH is between 10.0 and about 11.0.

12. A composition for the enzymatic treatment of cellulosic fibers of fabrics, comprising a neutral and/or alkaline cellulose whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type fungus of thegenus Chrysosporium, said composition further comprising one or more components that enhance the performance of the cellulase, wherein the components are selected from the group consisting of added proteases, detergents, and surfactants.

13. The composition of claim 12, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenlandicum, and Chrysosporium tropicum.

14. The composition of claim 13, wherein the fungus is Chrysosporium luckowense.

15. The composition of claim 12, wherein the cellulase is isolated or obtained from a wild-type fungus of the genus Chrysosporium.

16. The composition of claim 13, wherein the cellulase is isolated or obtained from a wild-type selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenlandicum, and Chrysosporium tropicum.

17. The composition of claim 14, wherein the cellulase is isolated or obtained from a wild-type fungus Chrysosporium lucknowense.

18. A composition for the enzymatic treatment of cellulosic fibers or fabrics, having from about 33 to 191 units of endo-1,4-β

-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type fungus of the genus Chrysosporium.

19. The composition of claim 18, wherein the fungus is selected from the group consisting of Chrysosporium luckowense, Chrysosporium pannorum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum.

20. The composition of claim 19, wherein the fungus is Chrysosporium lucknowense.

21. A composition for the enzymatic treatment of cellulosic fibers or fabrics, having from about 33 to 191 units of endo-1,4-β

-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose isolated or obtained from a wild-type fungus of the genus Chrysosporium.

22. The composition of claim 21, wherein the fungus is selected from the group consisting of Chrysosporium luckowense, Chrysosporium pannorum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum.

23. The composition of claim 22, wherein the fungus is Chrysosporium lucknowense.

24. A composition for the enzymatic treatment of cellulosic fibers or fabrics, having about 120 to 191 units of endo-1,4-β

-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type fungus of the genus Chrysosporium.

25. The composition of claim 24, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium lobatum, Chrysosporium merdarium, and Chrysosporium tropicum.

26. The composition of claim 25, wherein the fungus is Chrysosporium luckowense.

27. A composition for the enzymatic treatment of cellulose fibers or fabrics, having about 120 to 191 units of endo-1,4-β

-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose isolated or obtained from a wild-type fungus of the genus Chrysosporium.

28. The composition of claim 27, wherein the fungus is selected from the group consisting of Chrysosporium luckowense, Chrysosporium lobatum, Chrysosporium merdarium, and Chrysosporium tropicum.

29. The composition of claim 28, wherein the fungus is Chrysosporium luckowense.

30. A laundry detergent composition, comprising a cellulase whose amino acid sequence is encoded by a nucleic aid sequence from a wild-type fungus of the genus Chrysosporium, further comprising a surfactant.

31. The composition of claim 30, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum.

32. The composition of claim 31, wherein the fungus is Chrysosporium lucknownense.

33. A laundry detergent composition, comprising a cellulase isolated or obtained from a wild-type fungus of the genus Chrysosporium, further comprising a surfactant.

34. The composition of claim 33, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium, keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum.

35. The composition of claim 34, wherein the fungus is Chrysosporium luckowense.

36. A neutral/alkaline cellulase composition obtained from a wild-type fungus of the species Chrysosporium luckowense.

37. A neutral/alkaline cellulase composition obtained from a wild-type fungus of the species Chrysosporium lucknowense Garg 27K, accession number VKM F-3500D.

38. A neutral/alkaline cellulase composition obtained from a wild-type fungus of the genus Chrysosporium, with the proviso that the fungus is not Chrysosporium pruinosum.

39. A composition for the enzymatic treatment of cellulosic fibers or fabrics, having from 314 to 1537 units of endo-1,4-β

-glucanase activity per gram of protein, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose whose amino acid sequence is encodes by a nucleic acid sequence from a wild-type fungus of the genus Chrysosporium.

40. The composition of claim 39, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium lobatum, Chrysosporium merdarium, and Chrysosporium tropicum.

41. The composition of claim 40, wherein the fungus is Chrysosporium luckowense.

42. A composition for the enzymatic treatment of cellulosic fibers or fabrics, having from 314 to 1537 units of endo-1,4-β

-glucanase activity per gram of protein, as measured by an endoviscometric assay, of a neutral and/or alkaline cellulose isolated or obtained from a wild-type fungus of the genus Chrysosporium.

43. The composition of claim 42, wherein the fungus is selected from the group consisting of Chrysosporium luckowense, Chrysosporium lobatum, Chrysosporium merdarium, and Chrysosporium tropicum.

44. The composition of claim 43, wherein the fungus is Chrysosporium luckowense.