Biosensor-aided high-throughput screening of hyper-producing cells for malonyl-CoA-derived products

Microbial Cell Factories

Research

Jian-Ming Jin¹  Shuang-Yan Tang²  Ruinan Jin²  Wei Chen²  Heng Li³ 

[1] Beijing Key Laboratory of Plant Resources Research and Development, Beijing Technology and Business University, Beijing, China;CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China;CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China;University of Chinese Academy of Sciences, Beijing, China;

关键词: Whole-cell biosensor;    Malonyl-CoA;    High-throughput screening;    Triacetic acid lactone;    Phloroglucinol;   

DOI  :  10.1186/s12934-017-0794-6

 received in 2017-04-18, accepted in 2017-10-24,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundMalonyl-coenzyme A (CoA) is an important biosynthetic precursor in vivo. Although Escherichia coli is a useful organism for biosynthetic applications, its malonyl-CoA level is too low.ResultsTo identify strains with the best potential for enhanced malonyl-CoA production, we developed a whole-cell biosensor for rapidly reporting intracellular malonyl-CoA concentrations. The biosensor was successfully applied as a high-throughput screening tool for identifying targets at a genome-wide scale that could be critical for improving the malonyl-CoA biosynthesis in vivo. The mutant strains selected synthesized significantly higher titers of the type III polyketide triacetic acid lactone (TAL), phloroglucinol, and free fatty acids compared to the wild-type strain, using malonyl-CoA as a precursor.ConclusionThese results validated this novel whole-cell biosensor as a rapid and sensitive malonyl-CoA high-throughput screening tool. Further analysis of the mutant strains showed that the iron ion concentration is closely related to the intracellular malonyl-CoA biosynthesis.

【 授权许可】

CC BY   
© The Author(s) 2017