US2009163400A1PendingUtilityA1

Subtilase Variants

Assignee: NOVOZYMES ASPriority: May 5, 1995Filed: Aug 5, 2008Published: Jun 25, 2009

Est. expiryMay 5, 2015(expired)· nominal 20-yr term from priority

Inventors:Laurens Nicolaas Sierkstra Jan Klugkist Peter Markvardsen Claus Von Der Osten Peter Bauditz

C12N 9/54C11D 3/386

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References

Claims

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.

Claims

exact text as granted — not AI-modified

1 - 87 . (canceled) 
     
     
         88 . A modified subtilase comprising one or more of the following substitutions:
 (a) a substitution of the amino acid residue at position 167 with Met, Pro, or Trp,   (b) a substitution of the amino acid residue at position 170 with Ile, Met, or Val,   (c) a substitution of the amino acid residue at position 171 with Met, Pro or Trp, and   (d) a substitution of the amino acid residue at position 194 with Ile, Met, Phe, Trp, or Val,   
       wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         89 . The modified subtilase of  claim 88 , which comprises a substitution of the amino acid residue at position 167 with Met, Pro, or Trp. 
     
     
         90 . The modified subtilase of  claim 88 , which comprises a substitution of the amino acid residue at position 170 with Ile, Met, or Val. 
     
     
         91 . The modified subtilase of  claim 88 , which comprises a substitution of the amino acid residue at position 171 with Met, Pro or Trp. 
     
     
         92 . The modified subtilase of  claim 88 , which comprises a substitution of the amino acid residue at position 194 with Ile, Met, Phe, Trp, or Val. 
     
     
         93 . The modified subtilase of  claim 88 , 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. 
     
     
         94 . The modified subtilase of  claim 93 , wherein at least one further mutation is selected from the group consisting of K27R, *36D, S57P, N76D, G97N, S101G, V104A, V104N, V104Y, H120D, N123S, Q206E, N218S, M222A, M222S, T224S, K235L, and T274A. 
     
     
         95 . A detergent composition comprising a modified subtilase of  claim 88  and a surfactant. 
     
     
         96 . A modified subtilisin 309 comprising one or both of the following substitutions:
 (a) a substitution of Glu at position 136 with Ala, Asn, Cys, Gly, His, Ser, Thr, or Tyr, and   (b) a substitution at Y171A, Y171C, Y171G, Y171H, Y171I, Y171L, Y171M, Y171N, Y171P, Y171Q, Y171S, or Y171W,   
       wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         97 . The modified subtilisin 309 of  claim 96 , which comprises a substitution of Glu at position 136 with Ala, Asn, Cys, Gly, His, Ser, Thr, or Tyr. 
     
     
         98 . The modified subtilisin 309 of  claim 96 , which comprises Y171A, Y171C, Y171G, Y171H, Y171I, Y171L, Y171M, Y171N, Y171P, Y171Q, Y171S, or Y171W. 
     
     
         99 . The modified subtilisin 309 of  claim 96 , further comprising at least one further mutation at one or more of positions: 27, 36, 57, 76, 97, 101, 104, 120, 123, 194, 206, 218, 222, 224, 235 and 274. 
     
     
         100 . The modified subtilisin 309 of  claim 99 , 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. 
     
     
         101 . A detergent composition comprising a modified subtilisin 309 of  claim 96  and a surfactant. 
     
     
         102 . A modified subtilisin 147 comprising one or both of the following substitutions:
 (a) a substitution of Glu at position 136 with Ala, Asn, Cys, Gly, His, Ser, Thr, or Tyr,   (b) a substitution at Y171A, Y171C, Y171G, Y171H, Y171I, Y171L, Y171M, Y171N, Y171P, Y171Q, Y171S, or Y171W,   
       wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         103 . The modified subtilisin 147 of  claim 102 , further comprising at least one further mutation at one or more of positions: 27, 36, 57, 76, 97, 101, 104, 120, 123, 194, 206, 218, 222, 224, 235 and 274. 
     
     
         104 . The modified subtilisin 147 of  claim 103 , 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. 
     
     
         105 . A detergent composition comprising a modified subtilisin 147 of  claim 102  and a surfactant. 
     
     
         106 . A modified  Bacillus  protease PB92 comprising a substitution Y171A, Y171C, Y171G, Y171H, Y171I, Y171L, Y171M, Y171N, Y171P, Y171Q, Y171S, or Y171W, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         107 . The modified  Bacillus  protease PB92 of  claim 106 , further comprising at least one further mutation at one or more of positions: 27, 36, 57, 76, 97, 101, 104, 120, 123, 194, 206, 218, 222, 224, 235 and 274. 
     
     
         108 . The modified  Bacillus  protease PB92 of  claim 107 , 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. 
     
     
         109 . A detergent composition comprising a modified  Bacillus  protease PB92 of  claim 106  and a surfactant. 
     
     
         110 . A modified subtilisin BPN′ comprising a substitution of Lys at position 136 with Ala, Asn, Cys, Gly, His, Ser, Thr, or Tyr, wherein each position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         111 . The modified subtilisin BPN′ of  claim 110 , further comprising at least one further mutation at one or more of positions: 27, 36, 57, 76, 97, 101, 104, 120, 123, 194, 206, 218, 222, 224, 235 and 274. 
     
     
         112 . The modified subtilisin BPN′ of  claim 111 , 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. 
     
     
         113 . A detergent composition comprising a modified subtilisin BPN′ of  claim 111  and a surfactant. 
     
     
         114 . A modified subtilisin Carlsberg comprising a substitution of Lys at position 136 with Ala, Asn, Cys, Gly, His, Ser, Thr, or Tyr, wherein the position corresponds to a position of the amino acid sequence of subtilisin BPN′ (SEQ ID NO: 7). 
     
     
         115 . The modified subtilisin Carlsberg of  claim 114 , further comprising at least one further mutation at one or more of positions: 27, 36, 57, 76, 97, 101, 104, 120, 123, 194, 206, 218, 222, 224, 235 and 274. 
     
     
         116 . The modified subtilisin Carlsberg of  claim 115 , 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. 
     
     
         117 . A detergent composition comprising a modified subtilisin Carlsberg of  claim 114  and a surfactant.

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