Agrobacterium sp I-671 was used to production of D-HPG from 5-mono substituted hydantoin derivatives. The D-hydantoinase and carbamoylase involved in this biotransformation process were both strictly D-stereo specific. Their biosynthesis was found to be inducible by addition of uracil to the cultivation media. Permeabilization with toluene was necessary to convert the N-carbamoylhydroxyphenylglycine because of the low permeability of the cell membrane to the NCHPG. Permeabilized and purified carbamoylase required reducing agents such as DTT for enzyme activity. D-Hydantoinase and carbamoylase were purified through simple purification procedures including ammonium sulfate fractionation, DEAE-cellulose chromatograph and sephadex G-150 chromatograph, successively. The reverse reaction of D-Hydantoinase was neglisible compared with forward reaction rate. The carbamoylase activity was strongly inhibited by ammonium ions co-produced with D-HPG, whereas the hydantoinase activity under same conditions was not affected. Optimum temperature and pH were respectively 60℃ and 9 for the purified hydantoinase, 50℃ and 7.0 for the purified carbamoylase. The Km and Vm values were determined to be 1.28mM, 4,58mM/h for the D-hydantoinase and 3. 56mM, 6.30mM/h for carbamoylase, respectively. Zinc, nickel ions and p-chloromercurybenzoate inhibited the carbamoylase severely.
