Patent ID: 6440717
Filing Date: 2002-08-27
Classification: A61K,A61Q,C11D,C12N

Abstract:
A BPNâ€² variant having a modified amino acid sequence of the Subtilisin BPNâ€² wild-type amino acid sequence set forth in SEO ID NO:1, wherein the modified amino acid sequence comprises a substitution at one or more of positions 200, 201, 202, 203, 205, 206, 207, 208, 209, 210, 211, 212, 216, 219 or 220, whereina. when a substitution occurs at position 200, the substituting amino acid is His, Thr, Pro, Gly, Gln, Asn, Ser, Asp or Glu; b. when a substitution occurs at position 201, the substituting amino acid is Gly, Gln, Asn, Ser, Asp or Glu; c. when a substitution occurs at position 202, the substituting amino acid is Pro, Gln, Asn, Ser, Asp or Glu; d. when a substitution occurs at position 203, the substituting amino acid is Met, Cys, His, Pro, Gly, Gln, Asn, Ser, Asp or Glu; e. f. when a substitution occurs at position 205, the substituting amino acid is Leu, Met, Cys, Ala, His, Thr, Pro, Gly, Gln, Asn, Ser, Asp or Glu; f. when a substitution occurs at position 206, the substituting amino acid is Pro, Asn or Ser; g. when a substitution occurs at position 207, the substituting amino acid is Asp or Glu; h. when a substitution occurs at position 208, the substituting amino acid is Pro, Gly, Gln, Asn or Ser; wherein when position 208 is substituted, the variant is not a double mutation variant having a substitution at position 214, or a triple mutation variant having substitutions at positions 214 and 219; i. when a substitution occurs at position 209, the substituting amino acid is Ile, Val, Met, Cys, Ala, His, Thr, Pro, Gly, Gln, Asn, Ser, Asp or Glu; j. when a substitution occurs at position 210, the substituting amino acid is Gly, Gln, Asn, Ser, Asp or Glu; k. when a substitution occurs at position 211, the substituting amino acid is Ala, Pro, Gln, Asn, Ser, Asp or Glu; l. when a substitution occurs at position 212, the substituting amino acid is Gln, Ser, Asp or Glu; m. when a substitution occurs at position 216, the substituting amino acid is His, Thr, Pro, Gly, Gln, Asn, Ser, Asp or Glu; n. when a substitution occurs at position 219, the substituting amino acid is Pro, Gln, Asn, Ser, Asp or Glu; and o. when a substitution occurs at position 220, the substituting amino acid is Pro, Gly, Gln, Asn, Asp or Glu; whereby the BPNâ€² variant has decreased adsorption to, and increased hydrolysis of, an insoluble substrate as compared to wild-type subtilisin BPNâ€².