SummaryThe procedure for isolation of D-phenylglycine from its racemic mixture by enzymatic hydrolysis of L-enantiomer of N-acetyh,L-phenylglycine is described. For this hydrolysis, aminoacylase from Micrococcus agilis immobilized by sorption of DEAE-cellulose was applied. As is also shown, the course of enzymatic reaction can be directly controlled by spectrophotometric method.