Li Bingjuan, Li Yuxia, Bai Dongmei, Zhang Xin, Yang Huiying, Wang Jie, Liu Gang, Yue Juejie, Ling Yan, Zhou Dongsheng, Chen Huipeng
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing 100071, China.
Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
Sci Rep. 2014 Oct 24;4:6750. doi: 10.1038/srep06750.
Lactobacillus brevis alcohol dehydrogenase (Lb-ADH) catalyzes reduction of prochiral carbonyl compounds to chiral alcohol and meanwhile consumes its cofactor NADH into NAD(+), while the cofactor regeneration can be catalyzed by Candida boidinii formate dehydrogenase (Cb-FDH). This work presents three different Escherichia coli whole-cell biocatalyst systems expressing recombinant ADH/FDH, FDH-LIN1-ADH and FDH-LIN2-ADH, respectively, all of which display very high efficacies of prochiral carbonyl conversion with respect to conversion rates and enantiomeric excess values. ADH/FDH represents co-expression of Lb-ADH and Cb-FDH under different promoters in a single vector. Fusion of Lb-ADH and Cb-FDH by a linker peptide LIN1 (GGGGS)₂ or LIN2 (EAAAK)₂ generates the two bifunctional enzymes FDH-LIN1-ADH and FDH-LIN2-ADH, which enable efficient asymmetric reduction of prochiral ketones in whole-cell biotransformation.
短乳杆菌乙醇脱氢酶（Lb - ADH）催化前手性羰基化合物还原为手性醇，同时将其辅因子NADH消耗生成NAD(+)，而辅因子再生可由博伊丁假丝酵母甲酸脱氢酶（Cb - FDH）催化。这项工作展示了三种不同的大肠杆菌全细胞生物催化剂系统，分别表达重组ADH/FDH、FDH - LIN1 - ADH和FDH - LIN2 - ADH，所有这些系统在转化率和对映体过量值方面都表现出非常高的前手性羰基转化效率。ADH/FDH表示在单个载体中不同启动子下Lb - ADH和Cb - FDH的共表达。通过接头肽LIN1（GGGGS）₂或LIN2（EAAAK）₂将Lb - ADH和Cb - FDH融合产生两种双功能酶FDH - LIN1 - ADH和FDH - LIN2 - ADH，它们能够在全细胞生物转化中高效不对称还原前手性酮。
