Betaine drives efficient methyl supply in Escherichia coli From Metabolic Engineering

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Methylation reaction widely exists in animals, plants and microbial cells. Its reaction involves the methylation modification of DNA, proteins and many secondary metabolites including antibiotics. In most methylation reactions, S-adenosylmethionine (SAM) plays an important role as the only methylation donor. However, when methyl compounds are synthesized by exogenous methyltransferase or endogenous methyltransferase, the limited intracellular Sam concentration often limits the synthetic ability and efficiency of methyl compounds.

In order to solve the problem of insufficient intracellular methyl supply during the synthesis of methyl compounds and improve the methyl supply capacity and efficiency, Tao Yong group, Key Laboratory of microbial physiology and metabolic engineering, Institute of Microbiology, Chinese Academy of Sciences, recently published the research work entitled “improved methylation in E. coli via an effective methyl supply system driven by betaine” in metabolic engineering, an internationally renowned journal in the field of metabolic engineering and synthetic biology. Liu Qun, a direct doctoral student of the Institute of Microbiology of the Chinese Academy of Sciences, is the first author of the paper, and Lin Baixue project researcher and Tao Yong researcher of the Institute of Microbiology of the Chinese Academy of sciences are the corresponding authors.

In this study, betaine was used as a new methyl source. The exogenous betaine metabolism module was introduced into E. coli, and the exogenous module catalyzing the O-methylation of caffeic acid to form ferulic acid was used to characterize the methyl supply efficiency. 8.3 mm ferulic acid could be detected in the modified engineering bacteria for 24 hours, and the methyl supply capacity was increased by 12.6 times, which effectively improved the methyl supply capacity and efficiency. The research ideas include introducing the metabolic module of betaine to reconstruct the methionine cycle, improving the conversion efficiency of methionine to Sam, coupling exogenous adenosine to regenerate ATP system to make up for the cleavage of ATP molecules in the reaction process, blocking the SAH hydrolysis pathway of endogenous adenine, and primer the SAH hydrolysis pathway of exogenous adenosine to enrich adenosine. The reconstructed methionine cycle effectively improves the methyl supply capacity of the system. At the same time, the methyl transfer system can also be applied to methylation reactions catalyzed by different types of methyltransferases, such as the conversion of guanidine acetic acid to creatine catalyzed by N-methyltransferase, the conversion of protocatechuic acid to vanillic acid catalyzed by O-methyltransferase, and the methylation of DNA fragments in vitro, so as to improve its methyl transfer ability and efficiency.

This study is the first report to realize the efficient synthesis of methyl compounds by using betaine as a methyl source in E. coli. It is also the first report to introduce the metabolic module of betaine into heterologous host to improve methyl supply. This study provides an efficient methyl supply system with betaine as methyl source for the synthesis of methyl compounds, and lays a theoretical foundation for the study of intracellular methyl supply.

章链接: https://doi.org/10.1016/j.ymben.2022.02.004

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