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 I, 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.