A sensitive PnpR-based biosensor for p-nitrophenol detection.

A common aromatic and phenolic pollutant, p-nitrophenol (PNP), is widely used in various industry and has serious risk to the environmental health. Biosensors have been extensively employed as an alternative technology for pollutants monitoring. By mining the new sensing elements, more specific biosensors could be characterized for highly sensitive detection. Herein, the PnpR transcription factor was identified to activate the transcription of pnpC1 by binding to P promoter region in P. putida DLL-E4, and PNP was recognized as its specific inducer. The PnpR-based biosensor for detection of PNP was developed, demonstrating adequate sensitivity in a liquid solution with satisfactory specificity. The biosensor was optimized by adopting a transcriptional amplifier, which increased the maximum output by 149-fold, and improved the detection limit by 100-fold, from 1 mg/L to 10 μg/L. These biosensors had a linear range of 5-80 mg/L and 0.01-1.0 mg/L for PNP determination, respectively. Then, the agarose gel entrapment-based biosensor was constructed and allowed a good of PNP detection in the range of 5-60 mg/L in M9 solid agar within 70 min, and a detection sensitive of 16.8 mg/kg in soil. The good performance of the biosensor suggested its potential application of high-efficient and on-site detection in environmental matrices.