Directed Evolution of 4-Hydroxyphenylpyruvate Biosensors Based on a Dual Selection System

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 Feb 10

PMID 38338812

Authors

Hongxuan Du

Yaoyao Liang

Jianing Li

Xinyao Yuan

Fenglin Tao

Chengjie Dong

Zekai Shen

Guangchao Sui

Pengchao Wang

Affiliations

Soon will be listed here.

Abstract

Biosensors based on allosteric transcription factors have been widely used in synthetic biology. In this study, we utilized the ADP1 transcription factor PobR to develop a biosensor activating the P promoter when bound to its natural ligand, 4-hydroxybenzoic acid (4HB). To screen for PobR mutants responsive to 4-hydroxyphenylpyruvate(HPP), we developed a dual selection system in . The positive selection of this system was used to enrich PobR mutants that identified the required ligands. The following negative selection eliminated or weakened PobR mutants that still responded to 4HB. Directed evolution of the PobR library resulted in a variant where PobR was 5.1 times more reactive to 4-hydroxyphenylpyruvate than PobR. Overall, we developed an efficient dual selection system for directed evolution of biosensors.

Citing Articles

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Ding N, Yuan Z, Ma Z, Wu Y, Yin L Molecules. 2024; 29(15).

PMID: 39124917 PMC: 11313831. DOI: 10.3390/molecules29153512.

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