An enzymic process of production Of D- and L-phenylglycine is described. The unnatural amino acid DL-phenylglycine was synthesized from benzaldehyde and its DL-N-acetyl derivative obtained. Hog kidney acylase 1, an amidohydrolase, was used as catalyst to hydrolyse enantiospecifically with 100% conversion of L-N-acetylphenylglycine. L-Phenylglycine was obtained with 36% isolated yield, with over 99% of enantiomeric excess. After acid hydrolysis using 48% HBr (v/v), unreacted D-N-acetylphenylglycine, yielded D-phenylglycine in 26% isolated yield, with over 95% of enantiomeric excess. The present methodology constitutes a new preparative route to produce both enantiomers of phenylglycine and is suitable for industrial use. (c) 2005 Elsevier Ltd. All rights reserved.