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  • Title: Monitoring in vivo metabolic flux with a designed whole-cell metabolite biosensor of shikimic acid.
    Author: Li H, Liang C, Chen W, Jin JM, Tang SY, Tao Y.
    Journal: Biosens Bioelectron; 2017 Dec 15; 98():457-465. PubMed ID: 28715793.
    Abstract:
    Knowledge of intracellular metabolite levels is important for the understanding of metabolic flux distributions. Whole-cell biosensors of key metabolites are ideal for the monitoring of carbon flow in important metabolic pathways, thus guiding metabolic engineering for microbial improvement. However, lack of biosensors for metabolites of interests has limited their applications. In this study, a genetically encoded whole-cell biosensor specifically responding to shikimic acid has been developed by screening a site-saturation mutagenesis library of the binding pocket of a uric acid-responsive regulatory protein. This biosensor has been successfully applied in analyzing and engineering metabolic flux in the shikimic acid pathway, through genome-wide screening of gene targets critical for the pathway flux, and by improving the specific activity of pathway key enzyme, AroG. This work demonstrates the feasibility of monitoring metabolic flux with the aid of whole-cell biosensors designed for key metabolites.
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