Phosphoenolpyruvic acid biosensor based on transcription factor and application of phosphoenolpyruvic acid biosensor

By constructing a phosphoenolpyruvate biosensor based on CbbR protein and a specific promoter, the problem of not being able to monitor PEP concentration in real time in existing technologies has been solved, achieving highly sensitive and wide dynamic range PEP concentration monitoring, which is suitable for metabolic regulation research and strain screening.

CN122256341APending Publication Date: 2026-06-23JIANGNAN UNIV +1
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Patent Information

Application Number
CN202610096157.1
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-01-23
Publication Date
2026-06-23

AI Technical Summary

Technical Problem

Existing technologies cannot achieve real-time and accurate monitoring of phosphoenolpyruvate (PEP) concentration. Traditional methods are complex to operate and cannot dynamically reflect changes in intracellular metabolites, and require the addition of additional chemical substances to regulate metabolic pathways.

Method used

We constructed a biosensor based on phosphoenolpyruvate and LysR-type transcription factor CbbR. Using CbbR protein, a specific promoter, and a fluorescent reporter gene, we characterized PEP concentration through fluorescence signals, avoiding interference from other intracellular metabolites. We optimized the promoter and RBS sequence to improve sensitivity and responsiveness.

Benefits of technology

It enables non-invasive real-time monitoring of instantaneous changes in intracellular PEP concentration, exhibiting high sensitivity and excellent linear response, with a dynamic range of approximately 6 times. It can accurately distinguish different PEP concentration changes and is suitable for metabolic flux monitoring and strain screening.

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Abstract

The invention relates to a phosphoenolpyruvic acid biosensor based on transcription factors and application thereof, and belongs to the technical field of biology. A natural response mechanism of phosphoenolpyruvic acid and a LysR type transcription factor CbbR is utilized to construct the phosphoenolpyruvic acid response biosensor, main elements of the phosphoenolpyruvic acid response biosensor comprise a CbbR protein, a CbbR protein specific promoter and a fluorescence reporter gene, the concentration of phosphoenolpyruvic acid is represented by using the intensity of a fluorescence signal, and the concentration of the phosphoenolpyruvic acid is determined by using the fluorescence signal intensity. RBS sequence screening, sigma factor regulation and control and promoter engineering are combined, so that the sensitivity and the dynamic range of the biosensor are remarkably improved, the biosensor shows excellent time resolution, the dynamic change of the intracellular phosphoenolpyruvic acid level can be monitored in real time, and the application prospect is wide. And a brand new thought and means are provided for construction and optimization of a biological dynamic monitoring tool.
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