Journal
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS
Volume 888, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.mrgentox.2023.503639
Keywords
Whole-cell biosensor; Pigment biosynthesis; DNA damage; Oxidative stress; Stress response
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The emergence of new compounds in the past decade requires a high-throughput screening method for toxicity assay. In this proof-of-concept study, stress-responsive whole-cell biosensors were used to evaluate the damages induced by toxic chemicals. Nine stress-responsive promoters were selected to assemble a set of blue indigoidine-based biosensors. Dose-dependent increase of visible blue signal was observed in response to mutagens, showing the potential of these biosensors for detecting DNA damage and oxidative stress.
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, respon-sive 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 biolumines-cent 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 im-provements can further improve the biosensing performance in future studies.
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