The pervasive presence of organophosphate pesticides (OPs), such as chlorpyrifos (CPF), in aquatic ecosystems underscores the urgent need for sensitive and reliable detection methods to safeguard environmental and public health. This study addressed the critical need for a novel biosensor capable of detecting CPF and its toxic metabolite, 3,5,6-trichloro-2-pyridinol (TCP), with high sensitivity and selectivity, suitable for field applications in environmental monitoring. The study engineered a whole-cell biosensor based on E. coli strains that utilize the ChpR transcriptional regulator and the vioABCE gene cluster, providing a distinct visual and colorimetric response to CPF and TCP. The biosensor's performance was optimized and evaluated across various water matrices, including freshwater, seawater, and soil leachate. The biosensor demonstrated high sensitivity with a broad linear detection range, achieving limits of detection (LODs) at 0.8 μM for CPF and 7.813 nM for TCP. The linear regression concentration ranges were 1.6-12.5 μM for CPF and 15.6-125 nM for TCP, aligning with environmental standard limits and ensuring the biosensor's effectiveness in real-world scenarios. This innovative biosensing approach offers a robust, user-friendly tool for on-site environmental monitoring, significantly mitigating OPs contamination and advancing current detection technologies to meet environmental protection standards.
Keywords: Chlorpyrifos Detection; Environmental Monitoring; Organophosphate Pesticides; Visual Signal Transduction; Whole-cell Biosensors.
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