Preparation of immobilized hydantoinase stabilized with divalent metal ions

摘要

An immobilised hydantoinase containing a divalent metal ion selected from Mn++, Co++, Fe++, Ni++ and Mg++ is produced that is useful for the preparation of a D(-)(optionally substituted phenyl)glycine or N-carbamoyl derivative thereof by hydrolysis of a 5-(optionally substituted phenyl)hydantoin in the absence of air. Immobilization is carried out in the presence of the metal iona nd the metal ion stabilizes the hydantoinase and renders it capable of repeated re-use. The divalent metal ion may also be added to a reaction mixture containing the immobilized hydantoinase to minimize reduction of hydantoinase activity during hydrolysis of hydantoin. Cells that produce hydantoinase may be immobilized within or on a support, or hydantoinase separated from cells can be adsorbed on a positively charged polymeric support such as an ion exchange resin or covalently bonded to a polymeric support. A cross-linking agent such as glutaraldehyde may be used in immobilization. A preferred hydantoinase producing microorganism is Bacillus brevis IFO 12333.