Alpha-Amylase Variants

Abstract

The invention relates to a variant of a parent Termamyl-like alpha-amylase, comprising mutations in two, three, four, five or six regions/positions. The variants have increased stability at high temperatures (relative to the parent). The invention also relates to a DNA construct comprising a DNA sequence encoding an alpha-amylase variant of the invention, a recombinant expression vector which carries a DNA construct of the invention, a cell which is transformed with a DNA construct of the invention, the use of an alpha-amylase variant of the invention for washing and/or dishwashing, textile desizing, starch liquefaction, a detergent additive comprising an alpha-amylase variant of the invention, a manual or automatic dishwashing detergent composition comprising an alpha-amylase variant of the invention, a method for generating a variant of a parent Termamyl-like alpha-amylase, which variant exhibits increased.

1 . A variant alpha-amylase, said variant has an amino acid sequence which is at least 80% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8, has alpha-amylase activity and comprises a substitution of lysine at position 376, using the numbering of SEQ ID NO:4. 
     
     
         2 . The variant of  claim 1 , wherein the variant has an amino acid sequence which is at least 85% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         3 . The variant of  claim 1 , wherein the variant has an amino acid sequence which is at least 90% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         4 . The variant of  claim 1 , wherein the variant has an amino acid sequence which is at least 95% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         5 . The variant of  claim 1 , wherein the variant has an amino acid sequence which is at least 97% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         6 . The variant of  claim 1 , wherein the variant has an amino acid sequence which is at least 99% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         7 . A variant alpha-amylase, said variant has an amino acid sequence which is at least 80% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8, has alpha-amylase activity and comprises a substitution of alanine at position 356, using the numbering of SEQ ID NO:4. 
     
     
         8 . The variant of  claim 7 , wherein the variant has an amino acid sequence which is at least 85% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         9 . The variant of  claim 7 , wherein the variant has an amino acid sequence which is at least 90% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         10 . The variant of  claim 7 , wherein the variant has an amino acid sequence which is at least 95% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         11 . The variant of  claim 7 , wherein the variant has an amino acid sequence which is at least 97% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         12 . The variant of  claim 7 , wherein the variant has an amino acid sequence which is at least 99% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         13 . A variant alpha-amylase, said variant has an amino acid sequence which is at least 80% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8, has alpha-amylase activity and comprises a substitution of phenylalanine at position 358, using the numbering of SEQ ID NO:4. 
     
     
         14 . The variant of  claim 13 , wherein the variant has an amino acid sequence which is at least 85% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         15 . The variant of  claim 13 , wherein the variant has an amino acid sequence which is at least 90% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         16 . The variant of  claim 13 , wherein the variant has an amino acid sequence which is at least 95% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         17 . The variant of  claim 13 , wherein the variant has an amino acid sequence which is at least 97% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8. 
     
     
         18 . The variant of  claim 13 , wherein the variant has an amino acid sequence which is at least 99% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 or 8.