Claims
- 1. A modified subtilisin comprising a mutation in an amino acid sequence of a subtilisin, wherein the mutation is a substitution of a tyrosine at position 167, numbered according to the amino acid sequence of the mature subtilisin BPN', with a threonine or tryptophan.
- 2. A modified subtilisin according to claim 1, wherein the mutation is Y167T.
- 3. A modified subtilisin according to claim 1, wherein the mutation is Y167W.
- 4. A detergent composition, comprising a modified subtilisin according to claim 1 and a surfactant.
- 5. A detergent composition comprising a surfactant and a modified subtilisin, wherein the modified subtilisin comprises a mutation in an amino acid sequence of a subtilisin at a position numbered according to the amino acid sequence of the mature subtilisin BPN', wherein the mutation is selected from the group consisting of:
- (a) a histidine at position 17 substituted by a glutamine,
- (b) a histidine at position 39 substituted by a serine,
- (c) a glutamic acid at position 54 substituted with an aspartic acid,
- (d) a histidine at position 120 substituted with an asparagine,
- (e) a tyrosine at position 167 substituted with a glutarnic acid,
- (f) a tyrosine at position 167 substituted with a phenylalanine,
- (g) a tyrosine at position 171 substituted with a phenylalanine,
- (h) a tyrosine at position 171 substituted with a valine,
- (i) a tyrosine at position 192 substituted with a glutamic acid,
- (j) a tyrosine at position 192 substituted with a phenylalanine,
- (k) a tyrosine at position 209 substituted with a phenylalanine,
- (l) a tyrosine at position 214 substituted with a phenylalanine,
- (m) a histidine at position 226 substituted with a serine, and
- (n) a tyrosine at position 263 substituted with a phenylalanine.
- 6. The detergent composition according to claim 5, wherein the mutation is H17Q.
- 7. The detergent composition according to claim 5, wherein the mutation is H39S.
- 8. The detergent composition according to claim 5, wherein the mutation is E54D.
- 9. The detergent composition according to claim 5, wherein the mutation is H120N.
- 10. The detergent composition according to claim 5, wherein the mutation is Y167E.
- 11. The detergent composition according to claim 5, wherein the mutation is Y167F.
- 12. The detergent composition according to claim 5, wherein the mutation is Y171F.
- 13. The detergent composition according to claim 5, wherein the mutation is Y171V.
- 14. The detergent composition according to claim 5, wherein the mutation is Y192E.
- 15. The detergent composition according to claim 5, wherein the mutation is Y192F.
- 16. The detergent composition according to claim 5, wherein the mutation is Y209F.
- 17. The detergent composition according to claim 5, wherein the mutation is Y214F.
- 18. The detergent composition according to claim 5, wherein the mutation is H226S.
- 19. The detergent composition according to claim 5, wherein the mutation is Y263F.
- 20. The detergem composition according to claim 5, wherein the modified subtilisin further comprises a second mutation in the amino acid sequence of the subtilisin, wherein the second mutation is selected from the group consisting of
- (a) a lysine at position 27 substituted with an arginine,
- (b) a tyrosine at position 91 substituted with a phenylalanine,
- (c) a lysine at position 94 substituted with an arginine,
- (d) a histidine at position 120 substituted with an aspartic acid,
- (e) a tyrosine at position 209 substituted with a leucine,
- (f) a tyrosine at position 214 substituted with a threonine,
- (g) a lysine at position 235 substituted with a leucine,
- (h) a lysine at position 235 substituted with an arginine,
- (i) a lysine at position 237 substituted with an arginine,
- (j) a lysine at position 251 substituted with a glutamic acid, and
- (k) a lysine at position 251 substituted with an asparagine,
- wherein the mutation recited in claim 5 and the second mutation are at different positions of the subtilisin.
- 21. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S.
- 22. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: E54D+Y91F+K94R.
- 23. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: E54D+Y91F+K94R+H120D.
- 24. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: E54D+Y91F+K94R+H120N.
- 25. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: Y167F+Y171V+Y192F+Y209F+Y214T.
- 26. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K235L+K237R+K251E+Y263F.
- 27. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K235L+K237R+K251N+Y263F.
- 28. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H226S+K235L+K237R+K251N+Y263F.
- 29. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H226S+K235L+K237R+K251E+Y263F.
- 30. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K235R+K237R+K251E+Y263F.
- 31. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K235R+K237R+K251N+Y263F.
- 32. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H226S+K235R+K237R+K251N+Y263F.
- 33. The detergent composition according to claim 20, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H226S+K235R+K237R+K251E+Y263F.
- 34. The detergent composition according to claim 22, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K27R+E54D+Y91F+K94R+Y167F+Y171V+Y192F+Y209F+Y214T+K235L+K237R+K251E+Y263F.
- 35. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amirto acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120D+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235L+K237R+K251N+Y263F.
- 36. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120N+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235L+K237R+K251E+Y263F.
- 37. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120N+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235L+K237R+K251N+Y263F.
- 38. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120D+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235L+K237R+K251E+Y263F.
- 39. The detergent composition according to claim 22, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K27R+E54D+Y91F+K94R+Y167F+Y171V+Y192F+Y209F+Y214T+K235R+K237R+K251E+Y263F.
- 40. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120D+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235R+K237R+K251N+Y263F.
- 41. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120N+Y167F+Y171V+Y192F+Y209+Y214T+H226S+K235R+K237R+K251E+Y263F.
- 42. The detergem composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120N+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235R+K237R+K251N+Y263F.
- 43. The detergent composition according to claim 21, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: H17Q+K27R+H39S+E54D+Y91F+K94R+H120D+Y167F+Y171V+Y192F+Y209F+Y214T+H226S+K235R+K237R+K251E+Y263F.
- 44. The detergent composition according to claim 22, wherein the modified subtilisin comprises the following set of mutations in the amino acid sequence of the subtilisin: K27R+E54D+Y91F+K94R+Y209F+Y214T+K235R+K237R+K251E+Y263F.
- 45. The detergent composition according to claim 39, wherein the modified subtilisin further comprises mutations of the subtilisin of the histidine residues at positions 17, 39, 120 and 226 to neutral amino acid residues.
- 46. The detergent composition according to claim 5; wherein the subtilisin is selected from the group consisting of subtilisin BPN', subtilisin amylosacchariticus, subtilisin 168, subtilisin mesentericopeptidase, subtilisin Carlsberg, subtilisin DY, subtilisin 309, subtilisin 147, subtilisin thermitase, Bacillus PB92 protease, protease TW7, protease TW3, proteinase K and aqualysin.
- 47. The detergent composition according to claim 46, wherein the subtilisin is subtilisin 309.
- 48. The detergent composition according to claim 46, wherein the subtilisin is subtilisin 147.
Priority Claims (2)
Number |
Date |
Country |
Kind |
90610077 |
Dec 1990 |
EPX |
|
9027836 |
Dec 1990 |
GBX |
|
Parent Case Info
This is a divisional application of application Ser. No. 07/811,502, filed Dec. 20, 1991, now U.S. Pat. No. 5,482,849, the contents of which are incorporated herein by reference in their entirety.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
RE34606 |
Estell et al. |
May 1994 |
|
Divisions (1)
|
Number |
Date |
Country |
Parent |
811502 |
Dec 1991 |
|