Enzyme for the production of long chain peracid

Abstract

The present invention provides methods and compositions comprising at least one perhydrolase enzyme for cleaning and other applications. In some embodiments, the present invention provides methods and compositions for generation of long chain peracids. Certain embodiments of the present invention find particular use in applications involving cleaning, bleaching and disinfecting.

We claim: 
     
       1. An isolated perhydrolase enzyme, comprising:
 an amino acid substitution at position 204; or 
 amino acid substitutions at positions 12 and 22, 12 and 154, 12 and 194, 154 and 194, or 154 and 196; 
 wherein said enzyme perhydrolyzes long chain acyl ester substrates of at least six carbon atoms,; the amino acid positions are positionally equivalent to those in M. smegmatis perhydrolase having the amino acid sequence of SEQ ID NO: 2; and wherein the amino acid sequence of said enzyme is at least 90% identical to the amino acid sequence of the wild type perhydrolase of M. smegmatis, as set forth in SEQ ID NO: 2. 
 
     
     
       2. The isolated perhydrolase enzyme of  claim 1 , wherein said enzyme produces long chain peracid in the presence of a long chain acyl ester substrate and peroxide. 
     
     
       3. The isolated perhydrolase enzyme of  claim 1 , wherein said long chain acyl ester substrate contains a chain of at least nine carbon atoms. 
     
     
       4. The isolated perhydrolase enzyme of  claim 1 , wherein said enzyme comprises at least one substitution at an amino acid position equivalent to a position in M. smegmatis perhydrolase comprising the amino acid sequence set forth in SEQ ID NO:2, wherein said at least one substitution is selected from positions 12, 22, 59, 153, 154, 194, 196, and 204. 
     
     
       5. The isolated perhydrolase enzyme of  claim 4 , wherein said enzyme comprises at least one of the following amino acid substitutions: a Gly, Pro or Gln at position 12, a Trp at position 22, a Pro at position 59, a Pro at position 153, a Thr, Ser, Val or Gin at position 154, a Gly at position 194, a Ser, Gln Val, Gly, Pro, Ile or His at position 196, a Tyr or Trp at position 204, or any combination thereof, wherein said amino acid positions are positionally equivalent to positions 12, 22, 59, 153, 154, 194, 196 and 204 in the M. smegmatis perhydrolase of SEQ ID NO:2. 
     
     
       6. The isolated perhydrolase enzyme of  claim 1 , wherein said enzyme comprises at least one of the following amino acid substitutions: anAla at position 154 and a Met at position 194, a Gly at position 154 and a Val at position 194, or a Gly at position 12 and a Met at position 194, wherein said amino acid positions are positionally equivalent to positions 12, 154 and 194 in the M. smegmatis perhydrolase of SEQ ID NO:2. 
     
     
       7. The isolated perhydrolase enzyme of  claim 1 , wherein said enzyme has a perhydrolysis to hydrolysis ratio of greater than 1. 
     
     
       8. The isolated perhydrolase enzyme of  claim 1 , wherein said enzyme has a peracid hydrolysis rate that is lower than the peracid hydrolysis rate of SEQ ID NO:2. 
     
     
       9. An isolated perhydrolase enzyme, wherein said enzyme hydrolyzes long chain acyl ester substrates of at least six carbon atoms, and wherein the amino acid sequence of said enzyme is at least 90% identical to the amino acid sequence of the wild type perhydrolase of M. smegmatis, as set forth in SEQ ID NO: 2. 
     
     
       10. The isolated perhydolase enzyme of  claim 9 , wherein said enzyme produces long chain peracid in the presence of a long chain acyl ester substrate and peroxide. 
     
     
       11. The isolated perhydrolase enzyme of  claim 9 , wherein said long chain acyl ester substrate contains a chain of at least nine carbon atoms. 
     
     
       12. A cleaning composition comprising the perhydrolase enzyme of  claim 1 . 
     
     
       13. The cleaning composition of  claim 12 , wherein said composition further comprises a long chain acyl ester substrate and a source of peroxide. 
     
     
       14. The cleaning composition of  claim 13 , wherein said composition further comprises a surfactant. 
     
     
       15. The cleaning composition of  claim 12 , wherein said composition is a laundry detergent. 
     
     
       16. A cleaning method comprising contacting a substrate in the presence of the cleaning composition of  claim 12  to clean said substrate.