Protease variants and compositions
First Claim
Patent Images
1. A modified subtilisin 309 comprising one or both of the following substitutions:
- (a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, or Y167W, and (b) R170A, R170C, R170F, R170G, R170H, R170I, R170L, R170M, R170N, R170P, R170S, R170T, R170V, or R170W, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′
.
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Abstract
The present invention relates to enzymes produced by mutating the genes for a number of subtilases and expressing the mutated genes in suitable hosts are presented. The enzymes exhibit improved stability and/or improved wash performance in any detergent in comparison to their wild type parent enzymes. The enzymes are well-suited for use in any detergent and for some in especially liquid or solid shaped detergent compositions.
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Citations
60 Claims
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1. A modified subtilisin 309 comprising one or both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, or Y167W, and (b) R170A, R170C, R170F, R170G, R170H, R170I, R170L, R170M, R170N, R170P, R170S, R170T, R170V, or R170W, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′
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2. The modified subtilisin 309 of claim 1, which comprises Y167A.
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3. The modified subtilisin 309 of claim 1, which comprises Y167C.
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4. The modified subtilisin 309 of claim 1, which comprises Y167G.
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5. The modified subtilisin 309 of claim 1, which comprises Y167H.
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6. The modified subtilisin 309 of claim 1, which comprises Y167I.
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7. The modified subtilisin 309 of claim 1, which comprises Y167L.
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8. The modified subtilisin 309 of claim 1, which comprises Y167M.
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9. The modified subtilisin 309 of claim 1, which comprises Y167N.
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10. The modified subtilisin 309 of claim 1, which comprises Y167P.
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11. The modified subtilisin 309 of claim 1, which comprises Y167Q.
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12. The modified subtilisin 309 of claim 1, which comprises Y167S.
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13. The modified subtilisin 309 of claim 1, which comprises Y167T.
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14. The modified subtilisin 309 of claim 1, which comprises Y167V.
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15. The modified subtilisin 309 of claim 1, which comprises Y167W.
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16. The modified subtilisin 309 of claim 1, which comprises R170A.
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17. The modified subtilisin 309 of claim 1, which comprises R170C.
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18. The modified subtilisin 309 of claim 1, which comprises R170F.
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19. The modified subtilisin 309 of claim 1, which comprises R170G.
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20. The modified subtilisin 309 of claim 1, which comprises R170H.
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21. The modified subtilisin 309 of claim 1, which comprises R170I.
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22. The modified subtilisin 309 of claim 1, which comprises R170L.
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23. The modified subtilisin 309 of claim 1, which comprises R170M.
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24. The modified subtilisin 309 of claim 1, which comprises R170N.
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25. The modified subtilisin 309 of claim 1, which comprises R170P.
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26. The modified subtilisin 309 of claim 1, which comprises R170Q.
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27. The modified subtilisin 309 of claim 1, which comprises R170S.
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28. The modified subtilisin 309 of claim 1, which comprises R170T.
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29. The modified subtilisin 309 of claim 1, further comprising at least one further mutation at one or more of positions:
- 27, 36, 57, 76, 97, 101, 104, 120, 123, 206, 218, 222, 224, 235 and 274.
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30. The modified subtilisin 309 of claim 31, wherein at least one further mutation is selected from the group consisting of K27R, *36D, S57P, N76D, G97N, S101G, V104A, V104N, V104Y, H120D, N123S, A194P, Q206E, N218S, M222A, M222S, T224S, K235L, and T274A.
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31. A detergent composition comprising a modified subtilisin 309 of claim 1 and a surfactant.
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32. The detergent composition of claim 32, wherein the composition is in solid powdered form.
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33. The detergent composition of claim 32, wherein the composition is in liquid form.
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34. An isolated nucleic acid encoding a modified subtilisin 309 of claim 1.
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35. A vector comprising a nucleic acid of claim 34.
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36. A microbial host cell comprising a vector of claim 35.
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37. A method for producing a modified subtilisin 309, which comprises
(a) culturing a microbial host cell of claim 36 under conditions conducive to the expression and secretion of the modified subtilisin 309, and (b) recovering the modified subtilisin 309.
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38. A modified Bacillus protease PB92 comprising one or both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y1671, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, or Y167W, and (b) R170A, R170C, R170F, R170G, R170H, R170L, R170N, R170P, R170S, R170T, R1 70W, or R170Y, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′
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39. The modified Bacillus protease PB92 of claim 38, further comprising at least one further mutation at one or more of positions:
- 27, 36, 57, 76, 97, 101, 104, 120, 123, 206, 218, 222, 224, 235 and 274.
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40. The modified Bacillus protease PB92 of claim 39, wherein at least one further mutation is selected from the group consisting of K27R, *36D, S57P, N76D, G97N, S101G, V104A, V104N, V104Y, H120D, N123S, A194P, Q206E, N218S, M222A, M222S, T224S, K235L, and T274A.
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41. A detergent composition comprising a modified Bacillus protease PB92 of claim 38 and a surfactant.
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42. The detergent composition of claim 41, wherein the composition is in solid powdered form.
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43. The detergent composition of claim 41, wherein the composition is in liquid form.
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44. An isolated nucleic acid encoding a modified Bacillus protease PB92 of claim 38.
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45. A vector comprising a nucleic acid of claim 44.
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46. A microbial host cell comprising a vector of claim 45.
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47. A method for producing a modified Bacillus protease PB92, which comprises
(a) culturing a microbial host cell of claim 46 under conditions conducive to the expression and secretion of the modified Bacillus protease PB92, and (b) recovering the modified Bacillus protease PB92.
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48. A modified subtilisin 147 comprising one or both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, or Y167W, and (b) R170A, R170C, R170F, R170G, R170H, R170I R170L, R170M, R170N, R170P, R170S, R170T, R170V, R170W, or R170Y, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′
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49. The modified subtilisin 147 of claim 48, further comprising at least one further mutation at one or more of positions:
- 27, 36, 57, 76, 97, 101, 104, 120, 123, 206, 218, 222, 224, 235 and 274.
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50. The modified subtilisin 147 of claim 49, wherein at least one further mutation is selected from the group consisting of K27R, *36D, S57P, N76D, G97N, S101G, V104A, V104N, V104Y, H120D, N123S, A194P, Q206E, N218S, M222A, M222S, T224S, K235L, and T274A.
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51. A detergent composition comprising a modified subtilisin 147 of claim 48 and a surfactant.
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52. The detergent composition of claim 51, wherein the composition is in solid powdered form.
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53. The detergent composition of claim 51, wherein the composition is in liquid form.
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54. An isolated nucleic acid encoding a modified subtilisin 147 of claim 48.
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55. A vector comprising a nucleic acid of claim 54.
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56. A microbial host cell comprising a vector of claim 55.
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57. A method for producing a modified subtilisin 147, which comprises
(a) culturing a microbial host cell of claim 56 under conditions conducive to the expression and secretion of the modified subtilisin 147, and (b) recovering the modified subtilisin 147.
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58. A modified subtilsin 309, comprising both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, Y167V, or Y167W, and (b) R170A, R170C, R170F, R170 G, R170H, R170I, R170L, R170M, R170N, R170P, R170Q, R170S, R170T, R170V, or R170W, wherein each position corresponds to a position of the amino acid sequence of subtilsin BPN′
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59. A modified Bacillus protease PB92 comprising both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, Y167V, or Y167W, and (b) R170A, R170C, R170F, R170 G, R170H, R170I, R170L, R170M, R170N, R170P, R170Q, R170S, R170T, R170V, or R170W, wherein each position corresponds to a position of the amino acid sequence of subtilsin BPN′
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60. A modified subtilsin 147, comprising both of the following substitutions:
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(a) Y167A, Y167C, Y167G, Y167H, Y167I, Y167L, Y167M, Y167N, Y167P, Y167Q, Y167S, Y167T, Y167V, or Y167W, and (b) R170A, R170C, R170F, R170 G, R170H, R170I, R170L, R170M, R170N, R170P, R170Q, R170S, R170T, R170V, or R170W, wherein each position corresponds to a position of the amino acid sequence of subtilsin BPN′
.
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Specification