A panel of visual bacterial biosensors for the rapid detection of genotoxic and oxidative damage: A proof of concept study.

Chang-Ye Hui, Shun-Yu Hu, Xue-Qin Yang, Yan Guo

Mutation research. Genetic toxicology and environmental mutagenesis 2023 May-Jun

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The emergence of new compounds during the past decade requires a high-throughput screening method for toxicity assay. The stress-responsive whole-cell biosensor is a powerful tool to evaluate direct or indirect damages of biological macromolecules induced by toxic chemicals. In this proof-of-concept study, nine well-characterized stress-responsive promoters were first selected to assemble a set of blue indigoidine-based biosensors. The PuspA-based, PfabA-based, and PgrpE-based biosensors were eliminated due to their high background. A dose-dependent increase of visible blue signal was observed in PrecA-, PkatG-, and PuvrA-based biosensors, responsive to potent mutagens, including mitomycin and nalidixic acid, but not to genotoxic lead and cadmium. The PrecA, PkatG, and Ppgi gene promoters were further fused to a purple deoxyviolacein synthetic enzyme cluster. Although high basal production of deoxyviolacein is unavoidable, an enhanced visible purple signal in response to mitomycin and nalidixic acid was observed as dose-dependent, especially in PkatG-based biosensors. The study shows that a set of stress-responsive biosensors employing visible pigment as a reporter is pre-validating in detecting extensive DNA damage and intense oxidative stress. Unlike widely-used fluorescent and bioluminescent biosensors, the visual pigment-based biosensor can become a novel, low-cost, mini-equipment, and high-throughput colorimetric device for the toxicity assessment of chemicals. However, combining multiple improvements can further improve the biosensing performance in future studies. Copyright © 2023 Elsevier B.V. All rights reserved.

Citation

Chang-Ye Hui, Shun-Yu Hu, Xue-Qin Yang, Yan Guo. A panel of visual bacterial biosensors for the rapid detection of genotoxic and oxidative damage: A proof of concept study. Mutation research. Genetic toxicology and environmental mutagenesis. 2023 May-Jun;888:503639