New Intracellular Shikimic Acid Biosensor for Monitoring Shikimate Synthesis in Corynebacterium glutamicum

Area of Science:

• Metabolic Engineering
• Synthetic Biology
• Microbial Biotechnology

Background:

• Quantitative monitoring of intracellular metabolites using in vivo biosensors is crucial for strain identification and real-time product accumulation observation.
• Shikimic acid (SA) is a valuable precursor in various industrial applications, necessitating efficient production methods.

Purpose of the Study:

• To construct and characterize a novel shikimic acid (SA) biosensor for intracellular monitoring in Corynebacterium glutamicum.
• To utilize the SA biosensor for strain screening, high-throughput screening strategy development, and whole-cell biosensor engineering.

Main Methods:

• Construction of a shikimic acid (SA) biosensor utilizing the LysR-type transcriptional regulator (ShiR) from Corynebacterium glutamicum.
• Characterization of the SA biosensor's response to intracellular SA concentration at the single-cell level.
• Application of the biosensor for monitoring SA production, developing a ribosome binding site screening strategy, and engineering a whole-cell biosensor.

Main Results:

• The developed SA biosensor exhibited specific responses to intracellular SA concentration within a linear range of 19.5 ± 3.6 to 120.9 ± 1.2 fmole.
• The biosensor successfully monitored SA production in different C. glutamicum strains.
• A novel high-throughput screening strategy for ribosome binding sites was developed, leading to the engineering of high-yield SA-producing strains.

Conclusions:

• The novel intracellular SA biosensor is an effective tool for real-time monitoring of microbial SA production.
• This biosensor facilitates the rapid development and optimization of microbial strains for shikimic acid production.
• The study demonstrates the utility of biosensors in advancing metabolic engineering and microbial strain development.