Patent ID: 7358071

Claim:
A process for hydrolyzing a 3-hydroxynitrile to a 3-hydroxycarboxylic acid comprising the steps of: (a) contacting a 3-hydroxynitrile in an aqueous reaction mixture with an improved nitrilase catalyst characterized by at least a 1.8-fold improvement in nitrilase activity relative to the activity of the A. facilis 72W (ATCC 55746) nitrilase under identical reaction conditions, the improved nitrilase catalyst encoded by an isolated nucleic acid fragment; (i) encoding an enzymatically-active polypeptide having the amino acid sequence of SEQ ID NO:4; said isolated nucleic acid molecule further comprising at least one mutation that results in at least one amino acid substitution of SEQ ID NO:4, selected from the group consisting of: a) a substitution at position 210 with alanine, isoleucine, or cysteine; b) a substitution at position 65 with cysteine; c) a substitution at position 168 with lysine, valine or leucine; and d) a substitution at position 174 with isoleucine; (ii) having a nucleic acid sequence selected from the group consisting of SEQ ID NOs:5, 7, 11, 13, 15, and 17; or (iii) encoding an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 8, 12, 14, 16 and 18; and (b) recovering the 3-hydroxycarboxylic acid produced in step (a).