Protein engineering and iterative multimodule optimization for vitamin B6 production in Escherichia coli

Linxia Liu, Jinlong Li, Yuanming Gai, Zhizhong Tian, Yanyan Wang, Tenghe Wang, Pi Liu, Qianqian Yuan, Hongwu Ma, Sang Yup Lee () and Dawei Zhang ()
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Linxia Liu: Chinese Academy of Sciences
Jinlong Li: Chinese Academy of Sciences
Yuanming Gai: Chinese Academy of Sciences
Zhizhong Tian: Chinese Academy of Sciences
Yanyan Wang: Chinese Academy of Sciences
Tenghe Wang: Chinese Academy of Sciences
Pi Liu: Chinese Academy of Sciences
Qianqian Yuan: Chinese Academy of Sciences
Hongwu Ma: Chinese Academy of Sciences
Sang Yup Lee: Korea Advanced Institute of Science and Technology (KAIST)
Dawei Zhang: Chinese Academy of Sciences

Nature Communications, 2023, vol. 14, issue 1, 1-16

Abstract: Abstract Vitamin B6 is an essential nutrient with extensive applications in the medicine, food, animal feed, and cosmetics industries. Pyridoxine (PN), the most common commercial form of vitamin B6, is currently chemically synthesized using expensive and toxic chemicals. However, the low catalytic efficiencies of natural enzymes and the tight regulation of the metabolic pathway have hindered PN production by the microbial fermentation process. Here, we report an engineered Escherichia coli strain for PN production. Parallel pathway engineering is performed to decouple PN production and cell growth. Further, protein engineering is rationally designed including the inefficient enzymes PdxA, PdxJ, and the initial enzymes Epd and Dxs. By the iterative multimodule optimization strategy, the final strain produces 1.4 g/L of PN with productivity of 29.16 mg/L/h by fed-batch fermentation. The strategies reported here will be useful for developing microbial strains for the production of vitamins and other bioproducts having inherently low metabolic fluxes.

Date: 2023
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DOI: 10.1038/s41467-023-40928-0

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