高等学校化学学报

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (8): 1560.

• 研究论文 • 上一篇    下一篇

固定化细胞酶法拆分N-乙酰-D,L-3-甲氧基丙氨酸

熊吉滨, 刘均忠, 刘茜, 焦庆才   

  1. 南京大学生命科学学院, 医药生物技术国家重点实验室, 南京 210093
  • 收稿日期:2009-11-19 出版日期:2010-08-10 发布日期:2010-08-10
  • 通讯作者: 焦庆才, 男, 博士, 教授, 主要从事氨基酸及手性药物中间体研究. E-mail: jiaoqc@yahoo.com.cn
  • 基金资助:

    国家技术创新基金(批准号: 02CJ-13-01-16)资助.

Resolution of N-Acetyl-D,L-3-methoxy-alanine by Immobilized Cells with Aminoacylase

XIONG Ji-Bin, LIU Jun-Zhong, LIU Qian, JIAO Qing-Cai*   

  1. State Key Laboratory of Pharmaceutical Biotechnology, Nanjing Univeristy, Nanjing 210093, China
  • Received:2009-11-19 Online:2010-08-10 Published:2010-08-10
  • Contact: JIAO Qing-Cai. E-mail: jiaoqc@yahoo.com.cn
  • Supported by:

    国家技术创新基金(批准号: 02CJ-13-01-16)资助.

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摘要： 利用氨基酰化酶固定化细胞酶法拆分了N-乙酰-D,L-3-甲氧基丙氨酸. 考察了温度、pH值、底物浓度、金属离子和拆分时间对酶促反应的影响. 确定了氨基酰化酶固定化细胞手性拆分N-乙酰-D,L-3-甲氧基丙氨酸的最佳工艺条件为pH=7.0, 反应温度50 ℃及底物浓度500 mmol/L. 10-4 mol/L的Co2+和Mg2+对氨基酰化酶有显著激活作用, Cu2+和Zn2+对酶促反应有明显抑制作用. 在最佳条件下, 氨基酰化酶固定化细胞对N-乙酰-L-3-甲氧基丙氨酸的摩尔转化率达96%.

关键词: 氨基酰化酶, D,L-3-甲氧基丙氨酸, 固定化细胞, 手性拆分

Abstract: As a pharmaceutical intermediate, L-serine plays an important role in organic synthesis and peptide synthesis. Also N-acetyl-D,L-3-methoxy-alanine is an intermediate in synthesis of L-serine from methyl α-bromo-β-methoxypropionate. In this thesis, a new method of resolution of N-acetyl-D,L-3-methoxy-alanine by immobilized cells with aminoacylase was studied. Several factors of this enzymatic reaction were investigated. The optimal temperature and pH are 50 ℃ and 7.0. In 10 mL reaction solution, the optimal concentration of N-acetyl-D,L-3-methoxy-alanine is 500 mmol/L and the amount of immobilized cells is 1 g. 10-4 mol/L of Co2+ and Mg2+ are benefit to aminoacylase, while Cu2+ and Zn2+ show clearly inhibitory effect. Under the optimal conditions, molar conversion rate of N-acetyl-L-3-methoxy-alanine is 96%.

Key words: Aminoacylase, D,L-3-Methoxy-alanine, Immobilized cell, Chiral resolution

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