US2005003986A1PendingUtilityA1

Useful mutations of bacterial alkaline protease

63
Assignee: NOVOZYMES ASPriority: Jan 7, 1988Filed: Jul 21, 2004Published: Jan 6, 2005
Est. expiryJan 7, 2008(expired)· nominal 20-yr term from priority
Inventors:Sven HastrupSven BrannerFanny NorrisSteffen PetersenLeif Norskov-LauridsenVilly Johannes JensenDorrit Aaslyng
C12N 9/54C11D 3/386
63
PatentIndex Score
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Cited by
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Claims

Abstract

The present invention relates to mutations of a subtilisin gene which result in changes in the chemical characteristics of subtilisin enzymes. Mutations at specific nucleic acids of the subtilisin gene result in amino acid substitutions and consequently, altered enzyme function. Some of these mutant enzymes exhibit physical properties advantageous to industrial applications, particularly in the detergent industry, providing subtilisin which is more stable to oxidation, possesses greater protease activity, and exhibits improved washability.

Claims

exact text as granted — not AI-modified
1 . A modified subtilisin 309, comprising a substitution at one or more of the following positions: 
 53, 67, 124, 128, 131, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 168, 169, 170, 172, 218, 219, and 222    wherein each position corresponds to the position of the amino acid sequence of the mature subtilisin BPN′ as depicted in Table I(c):    
     
     
         2 . The modified subtilisin of  claim 1 , which has augmented protease activity or improved washing ability.  
     
     
         3 . The modified subtilisin of  claim 1 , comprising a substitution at position 53.  
     
     
         4 . The modified subtilisin of  claim 1 , comprising a substitution at position 67.  
     
     
         5 . The modified subtilisin of  claim 1 , wherein the substitution is with aspartic acid or glutamic acid.  
     
     
         6 . The modified subtilisin of  claim 1 , comprising a substitution at position 124.  
     
     
         7 . The modified subtilisin of  claim 1 , comprising a substitution at position 128.  
     
     
         8 . The modified subtilisin of  claim 1 , comprising a substitution at position 131.  
     
     
         9 . The modified subtilisin of  claim 1 , comprising a substitution at position 154.  
     
     
         10 . The modified subtilisin of  claim 1 , comprising a substitution at position 155.  
     
     
         11 . The modified subtilisin of  claim 1 , comprising a substitution at position 156.  
     
     
         12 . The modified subtilisin of  claim 1 , comprising a substitution at position 157.  
     
     
         13 . The modified subtilisin of  claim 1 , comprising a substitution at position 158.  
     
     
         14 . The modified subtilisin of  claim 1 , comprising a substitution at position 159.  
     
     
         15 . The modified subtilisin of  claim 1 , comprising a substitution at position 160.  
     
     
         16 . The modified subtilisin of  claim 1 , comprising a substitution at position 161.  
     
     
         17 . The modified subtilisin of  claim 1 , comprising a substitution at position 162.  
     
     
         18 . The modified subtilisin of  claim 1 , comprising a substitution at position 163.  
     
     
         19 . The modified subtilisin of  claim 1 , comprising a substitution at position 164.  
     
     
         20 . The modified subtilisin of  claim 1 , comprising a substitution at position 165.  
     
     
         21 . The modified subtilisin of  claim 1 , comprising a substitution at position 166.  
     
     
         22 . The modified subtilisin of  claim 1 , comprising a substitution at position 168.  
     
     
         23 . The modified subtilisin of  claim 22 , wherein the substitution is with alanine.  
     
     
         24 . The modified subtilisin of  claim 1 , comprising a substitution at position 169.  
     
     
         25 . The modified subtilisin of  claim 1 , comprising a substitution at position 170.  
     
     
         26 . The modified subtilisin of  claim 25 , wherein the substitution is with tyrosine.  
     
     
         27 . The modified subtilisin of  claim 1 , comprising a substitution at position 172.  
     
     
         28 . The modified subtilisin of  claim 1 , comprising a substitution at position 218.  
     
     
         29 . The modified subtilisin of  claim 28 , wherein the substitution is with serine.  
     
     
         30 . The modified subtilisin of  claim 1 , comprising a substitution at position 219.  
     
     
         31 . The modified subtilisin of  claim 30 , wherein the substitution is with methionine.  
     
     
         32 . The modified subtilisin of  claim 1 , comprising a substitution at position 222.  
     
     
         33 . The modified subtilisin of  claim 32 , wherein the substitution is with cysteine or alanine.  
     
     
         34 . A detergent composition comprising a modified subtilisin of  claim 1  and a surfactant.  
     
     
         35 . A modified subtilisin 309, comprising an insertion at one or more of the following positions: 
   36 , 56, 159, 164, 165 and 166    wherein each position corresponds to the position of the amino acid sequence of the mature subtilisin BPN′ as depicted in Table I(c).    
     
     
         36 . The modified subtilisin of  claim 35 , which has augmented protease activity or improved washing ability.  
     
     
         37 . The modified subtilisin of  claim 35 , comprising an insertion at position 6.  
     
     
         38 . The modified subtilisin of  claim 35 , comprising an insertion at position 56.  
     
     
         39 . The modified subtilisin of  claim 35 , comprising an insertion at position 159.  
     
     
         40 . The modified subtilisin of  claim 35 , comprising an insertion at position 164.  
     
     
         41 . The modified subtilisin of  claim 35 , comprising an insertion at position 165.  
     
     
         42 . The modified subtilisin of  claim 35 , comprising an insertion at position 166.  
     
     
         43 . A detergent composition comprising a modified subtilisin of  claim 35  and a surfactant.

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