Patent ID: 7300775

Claim:
A method for stereoselectively producing an alpha-substituted carboxylic acid, the method comprising (a) providing an aldehyde or a ketone; (b) providing a cyanide-containing compound; (c) providing an ammonia-containing compound or a compound comprising an ammonium salt or an amine; (d) providing a composition comprising a nitrilase, wherein the nitrilase has an amino acid sequence consisting of SEQ ID NO:2 or SEQ ID NO:4, or an enzymatically active fragment thereof, wherein the fragment retains the enzymatic function of SEQ ID NO:2 or SEQ ID NO:4; (e) contacting the aldehyde or ketone of step (a) with a cyanide-containing compound of step (b) and an ammonia-containing compound or a compound comprising an ammonium salt or an amine of step (c) such that an amino nitrile or a cyanohydrin intermediate is produced; and (f) contacting the amino nitrile or cyanohydrin intermediate of step (e) with the composition of step (d) such that the nitrilase stereoselectively hydrolyzes the amino nitrile or cyanohydrin intermediate to produce an alpha-substituted carboxylic acid, wherein said α-substituted carboxylic acid has the following structure: wherein: R 1 and R 2 are each independently —H, substituted or unsubstituted alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclic, wherein said substituents are lower alkyl, hydroxy, alkoxy, mercapto, cycloalkyl, heterocyclic, aryl, heteroaryl, aryloxy, or halogen or optionally R 1 and R 2 are linked to cooperate to form a functional cyclic moiety, and E is —N(R x ) 2 or —OH, wherein each R x is each independently —H or a lower alkyl.