Production of N-carbamoyl-D-phenylalanine by hydantoinase method coupled with in situ product removal

徐晓滢 姚忠 马哲 刘辉 周华 韦萍

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Production of N-carbamoyl-D-phenylalanine by hydantoinase method coupled with in situ product removal

更新时间：2025-12-10

- Production of N-carbamoyl-D-phenylalanine by hydantoinase method coupled with in situ product removal
- CIESC Journal Issue 4, (2007)
- 作者机构：
  
  南京工业大学制药与生命科学学院
- 作者简介：
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- Online First：05 April 2007，
  
  Published：05 April 2007
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徐晓滢姚忠马哲刘辉周华韦萍. Production of N-carbamoyl-D-phenylalanine by hydantoinase method coupled with in situ product removal[J]. CIESC Journal, 2007, (4).
DOI：

徐晓滢姚忠马哲刘辉周华韦萍. Production of N-carbamoyl-D-phenylalanine by hydantoinase method coupled with in situ product removal[J]. CIESC Journal, 2007, (4). DOI：

摘要

利用自行筛选的海因酶高产菌株Burkholderia cepecia njut01发酵后，经过硫酸铵分级沉淀、phenyl sepharose FF、DEAE sepharose FF等纯化步骤，得到初步纯化的D-海因酶；利用碳二亚胺法建立了一种基于EAH sepharose 4B的D-海因酶共价固定化方法，酶活回收率达到79.44%，固定化酶经100d 25个批次转化后仍可保持63.2％的初始酶活。将固定化酶填充为固定床反应器，考察了不同条件下固定床与固定床耦合离子交换原位分离（in situ product removal，ISPR）两种反应体系在转化过程中pH值变化及转化率的差异。结果表明，耦合离子交换原位分离技术可大幅度提高D，L-苄基海因转化率，经24h转化，N-氨甲酰-D-苯丙氨酸转化收率最高达62.725%，较采用单一固定床酶转化体系的转化率提高了89.3%。

Abstract

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.

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