期刊论文详细信息
Microbial Cell Factories
Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate l-pipecolic acid in Escherichia coli
Research
Hanxiao Ying1  Kequan Chen1  Jing Wang1  Pingkai Ouyang1  Sha Tao1  Xin Wang1  Weichao Ma1 
[1] State Key Laboratory of Materials Oriented Chemical Engineering, 211816, Nanjing, People’s Republic of China;College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 211816, Nanjing, People’s Republic of China;
关键词: Chiral intermediate biosynthesis;    Lysine cyclodeaminase;    -Pipecolic acid;    Metabolic engineering;    Cofactor engineering;   
DOI  :  10.1186/s12934-017-0666-0
 received in 2016-12-08, accepted in 2017-03-21,  发布年份 2017
来源: Springer
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【 摘 要 】

BackgroundThe six-carbon circular non-proteinogenic compound l-pipecolic acid is an important chiral drug intermediate with many applications in the pharmaceutical industry. In the present study, we developed a metabolically engineered strain of Escherichia coli for the overproduction of l-pipecolic acid from glucose.ResultsThe metabolic pathway from l-lysine to l-pipecolic acid was constructed initially by introducing lysine cyclodeaminase (LCD). Next, l-lysine metabolic flux from glucose was amplified by the plasmid-based overexpression of dapA, lysC, and lysA under the control of the strong trc promoter to increase the biosynthetic pool of the precursor l-lysine. Additionally, since the catalytic efficiency of the key enzyme LCD is limited by the cofactor NAD+, the intracellular pyridine nucleotide concentration was rebalanced by expressing the pntAB gene encoding the transhydrogenase, which elevated the proportion of LCD with bound NAD+ and enhanced l-pipecolic acid production significantly. Further, optimization of Fe2+ and surfactant in the fermentation process resulted in 5.33 g/L l-pipecolic acid, with a yield of 0.13 g/g of glucose via fed-batch cultivation.ConclusionsWe expanded the metabolic pathway for the synthesis of the chiral pharmaceutical intermediate l-pipecolic acid in E. coli. Using the engineered E. coli, a fast and efficient fermentative production of l-pipecolic acid was achieved. This strategy could be applied to the biosynthesis of other commercially and industrially important chiral compounds containing piperidine rings.

【 授权许可】

CC BY   
© The Author(s) 2017

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