Preparation of immobilized hydantoinase stabilized with divalent metal ions

A process for the production of a D(-) (unsubstituted or substituted phenyl) glycine or N-carbamoyl derivative thereof, which comprises:

• 1) providing a reusable, divalent metal ion-containing, immobilized enzyme preparation comprising a) cells of an organism producing an hydantoinase enzyme capable of hydrolyzing 5-(unsubstituted or substituted phenyl) hydantoin immobilized within or on a support or b) an hydantoinase enzyme capable of hydrolyzing 5-(unsubstituted or substituted phenyl) hydantoin absorbed on a positively charged polymeric support or covalently bonded to functional groups of a polymeric support and c) an amount effective to stabilize said immobilized enzyme preparation so that it is capable of reuse of a stabilizing divalent metal ion selected from the group consisting of Mn.sup.++, Co.sup.++, Fe.sup.++, Ni.sup.++ and Mg.sup.++ ;
• 2) hydrolyzing a 5-(unsubstituted or substituted phenyl) hydantoin in the absence of air by said reusable, divalent metal ion-containing, immobilized enzyme preparation to form a reaction product comprising D(-)(unsubstituted or substituted phenyl) glycine or N-carbamoyl derivative thereof;
• 3) separating said reusable, divalent metal ion-containing, immobilized enzyme preparation from said reaction product;
• 4) hydrolyzing fresh reactant comprising a 5-(unsubstituted or substituted phenyl) hydantoin in the absence of air by said reusable, divalent metal ion-containing, immobilized enzyme preparation recovered from step 3;
• and
• 5) repeating steps 2-4 for a plurality of cycles, whereby said recovered immobilized enzyme preparation is repeatedly reused to produce D(-)(unsubstituted or substituted phenyl) glycine or N-carbamoyl derivative thereof.