Thermostable D-hydantoinase from thermophilic Bacillus stearothermophilus SD-1 was used to produce N-carbamoyl-D-p-hydroxyphenylglycine (NC-HPG) from D, L-5-(4-hydroxyphenyl)/hydantoin (DL-5-HPH). Culture conditions for the production of the enzyme from B. stearothermophilus SD-1 were optimized. The D-hydantoinase was immobilized on various support matrices by adsorption, and DEAE-cellulose resin was Sound to be most effective ill terms of the activity recovery and the amount of protein bound. The activity of enzyme immobilized on DEAE-cellulose was retained >90%, and the optimal reaction conditions for the immobilized enzyme were determined to be 55 degrees C and pH 9.0, respectively. Immobilized enzyme was applied to the production of NC-HPG from DL-5-HPH in repealed hatch, and the production rate was maintained constantly over nine successive operations.