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›› 2005, Vol. 5 ›› Issue (6): 665-669.

• 生化工程专栏 • 上一篇    下一篇

生物催化不对称还原方法生产S-4-氯-3-羟基丁酸乙酯

钟萍,孙志浩   

  1. 江南大学生物工程学院生物催化研究室
  • 出版日期:2005-12-20 发布日期:2005-12-20

Production of Ethyl (S)-(-)-4-Chloro-3-hydroxybutyrate by Asymmetrical Reduction of Ethyl 4-Chloroacetoacetate with Aureobasidium pullulans SW0202

ZHONG Ping,SUN Zhi-hao   

  1. Laboratory of Biocatalysis, School of Biotechnology, Southern Yangtze University
  • Online:2005-12-20 Published:2005-12-20

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摘要： 通过筛选得到不对称还原4-氯乙酰乙酸乙酯(COBE)合成(S)-4-氯-3-羟基丁酸乙酯(S-CHBE)对映选择性好的微生物菌株出芽短梗霉菌(Aureobasidium pullulans)SW0202. 对其还原反应条件进行优化，得到最佳转化条件为：初始细胞浓度0.32 g/mL(以湿菌体计)，底物浓度20 g/L，pH 7.0，温度30℃. 在此优化条件下，产物浓度达20.32 g/L，摩尔转化率为79.6%，产物对映体过量值e.e.为97.8%. 分批添加底物可显著减小底物的抑制作用.

关键词: (S)-4-氯-3-羟基丁酸乙酯, 4-氯乙酰乙酸乙酯, 出芽短梗霉菌, 生物催化, 不对称还原反应

Abstract: Aureobasidium pullulans SW0202 with high enantioselectivity for asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to ethyl (S)-4-chloro-3-hydroxybutyrate (S-CHBE) was isolated. The asymmetric reduction of COBE to optically active CHBE catalyzed by Aureobasidium pullulans SW0202 in aqueous phase was also investigated. The reduction was conducted in K2HPO4-KH2PO4 buffer, and the concentrations of COBE and CHBE as well as optical purity of the product were determined by capillary gas chromatography (GC) and chiral capillary GC respectively. The results showed that the optimal conditions for the conversion were the initial cell concentration 0.32 g/mL(wet weight), the initial COBE concentration 20 g/L, pH 7.0 and the temperature 30℃. The molar conversion rate reached 79.6% (20.32 g/L CHBE), with 97.8% e.e. Subsequently the experimental results of batch supplement of substrate indicate that this strategy can greatly reduce substance inhibition.

Key words: ethyl (S)-(-)-4-chloro-3-hydroxybutyrate, ethyl 4-chloroacetoacetate, Aureobasidium pullulan, biocatalysis, asymmetric reduction