Bioretrosynthesis of Functionalized N-Heterocycles from Glucose via One-Pot Tandem Collaborations of Designed Microbes

Jing Feng; Ruifeng Li; Shasha Zhang; Yifan Bu; Yanchun Chen; Yinglu Cui; Baixue Lin; Yihua Chen; Yong Tao; Bian Wu

doi:10.1002/advs.202001188

Bioretrosynthesis of Functionalized N-Heterocycles from Glucose via One-Pot Tandem Collaborations of Designed Microbes

Adv Sci (Weinh). 2020 Jul 21;7(17):2001188. doi: 10.1002/advs.202001188. eCollection 2020 Sep.

Authors

Jing Feng^{1

2}, Ruifeng Li^{1

2}, Shasha Zhang^{1

2}, Yifan Bu^{1

2}, Yanchun Chen^{1

2}, Yinglu Cui¹, Baixue Lin¹, Yihua Chen¹, Yong Tao¹, Bian Wu¹

Affiliations

¹ CAS Key Laboratory of Microbial Physiological and Metabolic Engineering State Key Laboratory of Microbial Resources Institute of Microbiology Chinese Academy of Sciences Beijing 100101 P. R. China.
² University of Chinese Academy of Sciences Beijing China.

Abstract

The design of multistrain systems has markedly expanded the prospects of using long biosynthetic pathways to produce natural compounds. However, the cooperative use of artificially engineered microbes to synthesize xenobiotic chemicals from renewable carbohydrates is still in its infancy. Here, a microbial system is developed for the production of high-added-value N-heterocycles directly from glucose. Based on a retrosynthetic analysis, eleven genes are selected, systematically modulated, and overexpressed in three Escherichia coli strains to construct an artificial pathway to produce 5-methyl-2-pyrazinecarboxylic acid, a key intermediate in the production of the important pharmaceuticals Glipizide and Acipimox. Via one-pot tandem collaborations, the designed microbes remarkably realize high-level production of 5-methyl-2-pyrazinecarboxylic acid (6.2 ± 0.1 g L^-1) and its precursor 2,5-dimethylpyrazine (7.9 ± 0.7 g L^-1). This study is the first application of cooperative microbes for the total biosynthesis of functionalized N-heterocycles and provides new insight into integrating bioretrosynthetic principles with synthetic biology to perform complex syntheses.

Keywords: N‐heterocycles; bioretrosynthesis; cascade reactions; multistrain microbial systems; synthetic biology.