Patent EA-003295-B1 (EA)
Title: Cellulase of fungus of the genus chrysosporium and methods of use
Inventor: SOLOVJEVA IRINA VLADIMIROVNA
IPC: D06L4/40
Filed: 1997-09-30
Abstract: 1. A culture of Chrysosporium lucknowense Garg 27K having accession number VKM F-3500D producing a cellulase. 2. A composition having neutral and/or alkaline cellulase activity, obtained by a method which comprises growing a wild type or mutant fungus of the genus Chrysosporium lucknowense, wherein this cellulase has a peak of activity while the pH values are from nearly 5.5 to nearly 11.0 and at least 14 units of CMCase activity by 1 g of dry weight. 3. A composition according to claim 2 wherein the fungus is Chrysosporium lucknowense Garg 27K, accession number VKM F-3500D. 4. A composition according to claim 2 wherein the fungus is a mutant strain of Chrysosporium lucknowense Garg 27K. 5. A composition having neutral and/or alkaline cellulase activity having peak of cellulase activity having peak of cellulase activity while the pH values are from nearly 5.5 to nearly 11.0 and at least 14 units of CMCase activity by 1 g of dry weight obtained by a method which comprises growing mutant fungus of the genus Chrysosporium lucknowense in culture, in suitable medium. 6. A composition according to any one of claims 2-5 having cellulase activity at a temperature from about 40 degree C to about 60 degree C, at a pH from about 5.0 to about 12.0. 7. A composition according to any one of claims 2-5 having at least 50% of the optimal cellulase activity, at a pH from about 6.0 to about 7.0, at a temperature from about 40 degree C to about 60 degree C. 8. A composition according to any one of claims 2-5 wherein said cellulase activity is assayed by any one of the CMCase, RBBCMCase, endoviscometric or filter paper activity assays. 9. A substantially purified and isolated protein fraction, obtained from a composition according to claim 5, and having at least 50 % of its maximal cellulase activity at a pH between about 6.0 and about 7.0 as measured by any one of the CMCase, RBBCMCase, endoviscometric or titter paper activity assays. 10. An endoglucanase obtained from a fraction according to claim 9, having a molecular weight of about 60 kD and pI of about 3. 11. A cellobiohydrolase obtained from a fraction according to claim 10, having a molecular weight of about 60 kD and a pI of about 4. 12. A substantially purified and isolated neutral and/or alkaline cellulase enzyme, isolated from a protein fraction according to claim 10, and having a pI of between about 3 and about 5.5. 13. A cellulase according to claim 12 wherein said cellulase possesses either endoglucanase or cellobiohydrolase activity. 14. A cellulase according to claim 12 wherein said cellulase retains at least 50 % of its maximal cellulase activity at a pH between about 6.0 and about 7.0. 15. An endoglucanase obtained from a fraction according to claim 10 and having a molecular weight of about 25 kD. 16. An endoglucanase according to claim 15 having a value pI of about 4. 17. An endoglucanase obtained from a fraction according to claim 10 and having a molecular weight of about 70 kD. 18. An endoglucanase according to claim 17 having a value pI of about 4. 19. An endoglucanase obtained from a fraction according to claim 9 and having a molecular weight of about 43 kD. 20. An endoglucanase according to claim 19 having a value pI of about 3. 21. A detergent composition containing one or more purified enzymes isolated from a protein fraction according to claim 10-20, and further comprising a surfactant. 22. A fabric softening composition containing one or more purified enzymes obtained from the protein fraction according to claim 10-20. 23. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, comprising a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the Chrysosporium, said composition having a pH between about 8.0 and about l2.0. 24. The composition of claim 23, wherein the fungus is selected from the group consisting of Chrysosporiccm lucknowense, Chrysosporiccm pannorum, Chrysosporium pruinosccm, Chrysosporium keralinophilum, Chrysosporium lobatum; Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum. 25. The composition of claim 24, wherein the fungus is Chrysosporium lucknowense. 26. A composition according to any one of claims 23-25, wherein the cellulase is isolated or obtained from a wild-type or mutant fungus of the genus Chrysosporium. 27. A composition according to any one of claims 23-26, further comprising one or more components selected from the group consisting of pumice stones, abrasives, softeners, solvents, preservatives, bleaching agents, bluing agents, fluorescent dyes, antioxidants, solubilizers, detergents, surfactants, enzymes, builders, anti-redeposition agents, buffers, caking inhibitors, masking agents for factors inhibiting the cellulase activity, and cellulase activators. 28. The composition of claim 27, wherein the cellulase is isolated or obtained from a wild-type or mutant fungus of the genus Chrysosporium. 29. The composition of claim 23, wherein the pH is between 10.0 and about 11.0. 30. The composition of claim 24, wherein the pH is between 10.0 and about 11.0. 31. The composition of claim 25, wherein the pH is between 10.0 and about 11.0. 32. The composition of claim 26, wherein the pH is between 10.0 and about 11.0. 33. The composition of claim 27, wherein the pH is between 10.0 and about 11.0. 34. The composition of claim 28, wherein the pH is between 10.0 and about 11.0. 35. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, comprising a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus Chrysosporium, said composition further comprising one or more components selected from the group consisting of proteinases, detergents, and surfactants. 36. The composition of claim 35, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium kerarinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum. 37. The composition of claim 36, wherein the fungus is Chrysosporium lucknowense. 38. A composition as described in any one of claims 35-37, wherein the cellulase is isolated or obtained from a wild type or mutant fungus of the genus Chrysosporium. 39. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least 124 units of endo-1,4-beta-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus Chrysosporium. 40. The composition of claim 39, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobaxum, Chrysosporium merdarium, Chrysosporium queenslandieum, and Chrysosporium tropicum. 41. The composition of claim 40, wherein the fungus is Chrysosporium lucknowense. 42. A composition as described in any one of claims 39, wherein the cellulase is isolated or obtained from a wild type or mutant fungus of the genus Chrysosporium. 3. The composition of claim 42, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium keratinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslaruiicum, and Chrysosporium tropicum. 44. The composition of claim 43, wherein the fungus is Chrysosporium lucknowense. 45. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least 191 units of endo-1,4-beta-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus Chrysosporium. 46. The composition of claim 45, 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. 47. The composition of claim 46, wherein the fungus is Chrysosporium lucknowense. 48. A composition as described in any one of claims 45, wherein the cellulase is isolated or obtained from a wild type or mutant fungus of the genus Chrysosporium. 49. The composition of claim 49, 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. 50. The composition of claim 50, wherein the fungus is Chrysosporium lucknowense. 51. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 964 units of endo- 1,4-beta-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of a cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus Chrysosporium. 52. The composition of claim 51, wherein the fungus is selected from the group consisting of Chrysosporium lucknowense, Chrysosporium pannorum, Chrysosporium pruinosum, Chrysosporium kerazinophilum, Chrysosporium lobatum, Chrysosporium merdarium, Chrysosporium queenslandicum, and Chrysosporium tropicum. 53. The composition of claim 52, wherein the fungus is Chrysosporium lucknowense. 54. A composition as described in any one of claims 51, wherein the cellulase is isolated or obtained from a wild type or mutant fungus of the genus Chrysosporium. 55. The composition of claim 54, wherein the fungus is selected from