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A hybrid RNA-protein biosensor for high-throughput screening of adenosylcobalamin biosynthesis.
Yang, Xia; Wang, Huiying; Ding, Dongqin; Fang, Huan; Dong, Huina; Zhang, Dawei.
Afiliación
  • Yang X; College of Biological and Pharmaceutical Sciences, China Three Gorges University, Yichang, Hubei, 443002, China.
  • Wang H; Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin,300308, China.
  • Ding D; Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
  • Fang H; Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin,300308, China.
  • Dong H; Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
  • Zhang D; Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin,300308, China.
Synth Syst Biotechnol ; 9(3): 513-521, 2024 Sep.
Article en En | MEDLINE | ID: mdl-38680948
ABSTRACT
Genetically encoded circuits have been successfully utilized to assess and characterize target variants with desirable traits from large mutant libraries. Adenosylcobalamin is an essential coenzyme that is required in many intracellular physiological reactions and is widely used in the pharmaceutical and food industries. High-throughput screening techniques capable of detecting adenosylcobalamin productivity and selecting superior adenosylcobalamin biosynthesis strains are critical for the creation of an effective microbial cell factory for the production of adenosylcobalamin at an industrial level. In this study, we developed an RNA-protein hybrid biosensor whose input part was an endogenous RNA riboswitch to specifically respond to adenosylcobalamin, the inverter part was an orthogonal transcriptional repressor to obtain signal inversion, and the output part was a fluorescent protein to be easily detected. The hybrid biosensor could specifically and positively correlate adenosylcobalamin concentrations to green fluorescent protein expression levels in vivo. This study also improved the operating concentration and dynamic range of the hybrid biosensor by systematic optimization. An individual cell harboring the hybrid biosensor presented over 20-fold higher fluorescence intensity than the negative control. Then, using such a biosensor combined with fluorescence-activated cell sorting, we established a high-throughput screening platform for screening adenosylcobalamin overproducers. This study demonstrates that this platform has significant potential to quickly isolate high-productive strains to meet industrial demand and that the framework is acceptable for various metabolites.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Synth Syst Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: China
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Synth Syst Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: China