The purified D-hydantoinase was immobilized on EAH sepharose 4B via the carbodiimide method with a yield of enzyme activity up to 79.44%.The immobilized hydantoinase showed remarkable stability at 4℃.An integrated pr...The purified D-hydantoinase was immobilized on EAH sepharose 4B via the carbodiimide method with a yield of enzyme activity up to 79.44%.The immobilized hydantoinase showed remarkable stability at 4℃.An integrated process of N-carbamoyl-D-phenylalanine(N-D-Phe)synthesis from D,L-5-benzylhydantoin(D,L-BH)catalyzed by immobilized D-hydantoinase coupled with an ion-exchange unit for in situ product removal(ISPR)was established.The variation of pH and conversion in the fixed-bed reactor with or without ISPR was compared at different temperatures,initial substrate concentrations and volumes of adsorbent.Within 24 h,the pH value in the reactor with ISPR could be kept at the alkaline range,which was beneficial to the enzymatic conversion and racemization of L-5-benzyl hydantoinase.This led to a higher overall conversion of 62.725% under optimal operation conditions,an increase of 89.3% compared with the fixed-bed reactor without ISPR.展开更多
对BurkholderiacepeciaJS 02"一菌双酶法"制备对羟基 D 苯甘氨酸过程的调控进行了研究,探讨了反应液pH值对于酶反应各个进程的影响,实施了调节反应液pH恒定为7 2的调控策略,使得D 海因酶与D N 氨甲酰基氨基酸水解酶同时具有...对BurkholderiacepeciaJS 02"一菌双酶法"制备对羟基 D 苯甘氨酸过程的调控进行了研究,探讨了反应液pH值对于酶反应各个进程的影响,实施了调节反应液pH恒定为7 2的调控策略,使得D 海因酶与D N 氨甲酰基氨基酸水解酶同时具有较高的活力,并且较未调控前更有利于L 对羟基苯海因的消旋及DL 对羟基苯海因的溶解,底物转化率与产品收率分别由未调控时的92%与85%提高至99 5%与94%,反应时间由40h缩短至30h,生产速率由0 462g/(L·h)提高至0 681g/(L·h)展开更多
文摘The purified D-hydantoinase was immobilized on EAH sepharose 4B via the carbodiimide method with a yield of enzyme activity up to 79.44%.The immobilized hydantoinase showed remarkable stability at 4℃.An integrated process of N-carbamoyl-D-phenylalanine(N-D-Phe)synthesis from D,L-5-benzylhydantoin(D,L-BH)catalyzed by immobilized D-hydantoinase coupled with an ion-exchange unit for in situ product removal(ISPR)was established.The variation of pH and conversion in the fixed-bed reactor with or without ISPR was compared at different temperatures,initial substrate concentrations and volumes of adsorbent.Within 24 h,the pH value in the reactor with ISPR could be kept at the alkaline range,which was beneficial to the enzymatic conversion and racemization of L-5-benzyl hydantoinase.This led to a higher overall conversion of 62.725% under optimal operation conditions,an increase of 89.3% compared with the fixed-bed reactor without ISPR.
