Patent ID: 6576454
Filing Date: 2003-06-10
Classification: C07K,C11D,C12N,Y10S

Abstract:
A method of producing a modified subtilisin enzyme having esterase and amidase activities, said method comprising providing a modified enzyme wherein a cysteine residue is substituted for an amino acid residue selected from the group consisting of residues 62, 166, 217 and 222, wherein said amino acid residue is numbered according to its equivalent in the amino acid sequence of Bacillus amyloliquefaciens subtilisin BPNâ€² (SEQ ID NO: 2), and wherein said cysteine residue is modified by replacing the thiol hydrogen in the cysteine residue with a thiol side chain selected from the group consisting of â€”â€”SCH3, â€”â€”SCH2CH3, â€”â€”SCH2CH(CH3)2, â€”â€”S(CH2)4CH3, â€”S(CH2)5CH3, â€”â€”S(CH2)9 CH3â€”â€”SCH2 C6H5, â€”â€”SCH2, CH2, NH3+, â€”â€”SCH2, CH2, SO3âˆ’, â€”â€”SCH2, (p-COOHâ€”â€”C6H4,), and â€”â€”SCH 2,C6 F5 to form the modified enzyme.