1. Engineering Escherichia coli for high-yielding 2,5-Dimethylpyrazine synthesis from L-Threonine by reconstructing metabolic pathways and enhancing cofactors regeneration
- Author
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Xin-Xin Liu, Yao Wang, Jian-Hui Zhang, Yun-Feng Lu, Zi-Xing Dong, Chao Yue, Xian-Qing Huang, Si-Pu Zhang, Dan-Dan Li, Lun-Guang Yao, and Cun-Duo Tang
- Subjects
2,5-dimethylpyrazine ,Metabolic engineering ,Microbial cell factories ,Aminoacetone oxidase ,Threonine transporter ,Whole-cell catalysis ,Biotechnology ,TP248.13-248.65 ,Fuel ,TP315-360 - Abstract
Abstract 2,5-Dimethylpyrazine (2,5-DMP) is important pharmaceutical raw material and food flavoring agent. Recently, engineering microbes to produce 2,5-DMP has become an attractive alternative to chemical synthesis approach. In this study, metabolic engineering strategies were used to optimize the modified Escherichia coli BL21 (DE3) strain for efficient synthesis of 2,5-DMP using L-threonine dehydrogenase (EcTDH) from Escherichia coli BL21, NADH oxidase (EhNOX) from Enterococcus hirae, aminoacetone oxidase (ScAAO) from Streptococcus cristatus and L-threonine transporter protein (EcSstT) from Escherichia coli BL21, respectively. We further optimized the reaction conditions for synthesizing 2,5-DMP. In optimized conditions, the modified strain can convert L-threonine to obtain 2,5-DMP with a yield of 2897.30 mg/L. Therefore, the strategies used in this study contribute to the development of high-level cell factories for 2,5-DMP. Graphical Abstract
- Published
- 2024
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