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Fermentative production of enantiomerically pure S-1,2-propanediol from glucose by engineered E. coli strain
Lingfeng Zhu1,2, Xiangchen Guan1, Nengzhong Xie3
1CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
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.
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