Directed evolution of formate dehydrogenase and its application in the biosynthesis of L-phenylglycine from phenylglyoxylic acid

文献类型： 外文期刊

第一作者： Tang, Cun-Duo

作者： Tang, Cun-Duo;Zhang, Zhen-Hua;Shi, Hong-Ling;Xie, Yu-Li;Yang, Tian-Tian;Lu, Yun-Feng;Kan, Yun-Chao;Yao, Lun-Guang;Tang, Cun-Duo;Zhang, Zhen-Hua;Shi, Hong-Ling;Xie, Yu-Li;Yang, Tian-Tian;Lu, Yun-Feng;Kan, Yun-Chao;Yao, Lun-Guang;Tang, Cun-Duo;Zhang, Zhen-Hua;Shi, Hong-Ling;Xie, Yu-Li;Yang, Tian-Tian;Lu, Yun-Feng;Kan, Yun-Chao;Yao, Lun-Guang;Zhang, Si-Pu;Bai, Fang-Hui

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关键词： Formate dehydrogenase; L-phenylglycine; Directed evolution; Molecular docking; Biosynthesis

期刊名称：MOLECULAR CATALYSIS （ 影响因子：5.062； 五年影响因子：4.918 ）

ISSN： 2468-8231

年卷期： 2021 年 513 卷

页码：

收录情况： SCI

摘要： Formate dehydrogenase (FDH) is a D-2-hydroxy acid dehydrogenase, and catalyzes the oxidation of formate to carbon dioxide, coupled with reduction of NAD(+) to NADH that plays a key role in the process of NADH regeneration. In order to obtain high activity formate dehydrogenase mutants, the formate dehydrogenase CbFDHC23S was used as the parent to conduct two rounds of directed evolution, and a mutant M2 was obtained which specific activity was about 4 times more than the parent and was more suitable for coenzyme regeneration under physiological conditions. Then, the molecular mechanism of temperature characteristic and catalytic efficiency change was preliminarily elucidated by computer-aided method. Finally, an engineered E. coli strain was established to co-express formate dehydrogenase and L-leucine dehydrogenase and enantioselectively transform phenylglyoxylic acid to give L- phenylglycine (e.e. >99%), the yield and space-time yield of L- phenylglycine can reach 90.46% and 82.07 g.L-1.d(-1). This study laid a theoretical foundation for the green biosynthesis of food additives such as chiral alcohols and amino acid derivatives catalyzed by FDH coupling to enhance the regeneration capacity of NADH, reduce the regeneration cost of NADH, and achieve high efficiency and low cost.

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