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通过工程改造的大肠杆菌菌株从葡萄糖中发酵生产对映体纯的S-1,2-丙二醇。

Fermentative production of enantiomerically pure S-1,2-propanediol from glucose by engineered E. coli strain.

作者信息

Zhu Lingfeng, Guan Xiangchen, Xie Nengzhong, Wang Limin, Yu Bo, Ma Yanhe

机构信息

CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.

University of Chinese Academy of Sciences, Beijing, 100049, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2016 Feb;100(3):1241-1251. doi: 10.1007/s00253-015-7034-y. Epub 2015 Oct 10.

Abstract

The pure stereoisomers of 1,2-propanediol (1,2-PDO) could be used as starting materials to synthesize high value-added specialty chemicals and chiral pharmaceutical products. As the stereoisomers of 1,2-PDO cannot be obtained by traditional chemical synthesis processes, biotechnological processes have gained increasing attention. However, to our knowledge, the production of S-1,2-PDO directly from glucose has not been previously reported. In this study, we demonstrate a novel artificial pathway to convert L-lactic acid to S-1,2-PDO and its integration into the genome of Escherichia coli strain BW25113∆poxB with synchronous deletion of genes responsible for branch metabolic pathways from glucose. L-lactate production was increased by replacing the native D-lactate dehydrogenase with the L-lactate dehydrogenase from Bacillus coagulans. The methylglyoxal bypass pathway was blocked to avoid synthesis of a racemic mixture of D- and L-lactate and prevent the accumulation of methylglyoxal, a toxic intermediate. To further improve the yield of S-1,2-PDO, a novel cofactor regeneration system was introduced by combining pyruvate decarboxylase and acetaldehyde-CoA dehydrogenase II to simultaneously regenerate NADH and the CoA donor of acetyl-CoA for the lactate conversion pathway. Finally, 13.7 mM S-1,2-PDO with >99 % enantiomeric purity was directly produced from glucose by disrupting the major carbon-competing pathways and strengthening the lactate transformation pathway. This study demonstrates the first attempt to synthesize S-1,2-PDO by direct fermentation of glucose.

摘要

1,2 - 丙二醇(1,2 - PDO)的纯立体异构体可作为起始原料用于合成高附加值的特种化学品和手性药物产品。由于1,2 - PDO的立体异构体无法通过传统化学合成工艺获得,生物技术工艺受到了越来越多的关注。然而,据我们所知,此前尚未有关于直接从葡萄糖生产S - 1,2 - PDO的报道。在本研究中,我们展示了一条将L - 乳酸转化为S - 1,2 - PDO的新型人工途径,并将其整合到大肠杆菌BW25113∆poxB菌株的基因组中,同时删除负责葡萄糖分支代谢途径的基因。通过用凝结芽孢杆菌的L - 乳酸脱氢酶替代天然的D - 乳酸脱氢酶来提高L - 乳酸产量。阻断甲基乙二醛旁路途径以避免合成D - 乳酸和L - 乳酸的外消旋混合物,并防止有毒中间体甲基乙二醛的积累。为了进一步提高S - 1,2 - PDO的产量,引入了一种新型辅因子再生系统,该系统结合丙酮酸脱羧酶和乙醛 - CoA脱氢酶II,同时为乳酸转化途径再生NADH和乙酰 - CoA的CoA供体。最后,通过破坏主要的碳竞争途径并强化乳酸转化途径,直接从葡萄糖生产出了对映体纯度>99%的13.7 mM S - 1,2 - PDO。本研究展示了首次尝试通过葡萄糖直接发酵合成S - 1,2 - PDO。

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