Stereoselective reduction of ethyl 4-chloro-3-oxobutanoate by Escherichia coli transformant cells coexpressing the aldehyde reductase and glucose dehydrogenase genes
Stereoselective reduction of ethyl 4-chloro-3-oxobutanoate by Escherichia coli transformant cells coexpressing the aldehyde reductase and glucose dehydrogenase genes
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DOI：
10.1007/s002530051421
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发表时间：
1999-04-01
影响因子：
5
通讯作者：
Shimizu, S
中科院分区：
文献类型：
--
作者：
Kataoka, M;Yamamoto, K;Shimizu, S
以共表达沙门氏芽孢菌醛还原酶基因和巨芽孢杆菌葡萄糖脱氢酶基因的大肠杆菌细胞为催化剂，研究了4-氯-3-氧丁酸乙酯（COBE）不对称还原为(R)-4-氯-3-羟基丁酸乙酯[(R)-CHBE]。在有机溶剂-水两相体系中，有机相生成的(R)-CHBE为1610 mM (268 mg/ml)，摩尔产率为94.1%，光学纯度为91.7%。计算得到NADP（+）转化为CHBE的数量为13 500 mol/mol。由于使用携带pKAR和pACGD的大肠杆菌JM109细胞作为催化剂简单，不需要添加GDH或分离酶，因此对(R)-CHBE的实际合成非常有利。
The asymmetric reduction of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (R)-4-chloro-3-hydroxybutanoate [(R)-CHBE] using Escherichia coli cells, which coexpress both the aldehyde reductase gene from Sporobolomyces salmonicolor and the glucose dehydrogenase (GDH) gene from Bacillus megaterium as a catalyst was investigated. In an organic solvent-water two-phase system, (R)-CHBE formed in the organic phase amounted to 1610 mM (268 mg/ml), with a molar yield of 94.1% and an optical purity of 91.7% enantiomeric excess. The calculated turnover number of NADP(+) to CHBE formed was 13 500 mol/mol. Since the use of E. coli JM109 cells harboring pKAR and pACGD as a catalyst is simple, and does not require the addition of GDH or the isolation of the enzymes, it is highly advantageous for the practical synthesis of (R)-CHBE.