1. A highly sensitive biosensor with a single-copy evolved sensing cassette for chlorpyrifos pesticide detection.
- Author
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Chouichit P, Whangsuk W, Sallabhan R, Mongkolsuk S, and Loprasert S
- Subjects
- Bacterial Proteins genetics, Bacterial Proteins metabolism, Directed Molecular Evolution, Escherichia coli drug effects, Escherichia coli genetics, Escherichia coli metabolism, Glycine analogs & derivatives, Glycine metabolism, Limit of Detection, Piperazines pharmacology, Promoter Regions, Genetic, Recombinant Proteins genetics, Recombinant Proteins metabolism, Sinorhizobium meliloti genetics, Sulfatases genetics, Sulfatases metabolism, Transcription Factors genetics, Transcription Factors metabolism, Biosensing Techniques methods, Chlorpyrifos isolation & purification, Pesticides isolation & purification
- Abstract
A formylglycine-generating enzyme (FGE)-sulfatase-based whole-cell biosensor was genetically improved into a single-copy system by integrating the Sinorhizobium meliloti transcriptional activator ChpR and the chpA promoter-FGE-sulfatase fusion into the Escherichia coli chromosome. The sensitivity was further enhanced through a random mutagenesis of the chpR . The new integrated biosensor offered both a lower detection limit [5 nM chlorpyrifos (CPF)] and fluorescence background. The ready-to-use kit was developed using silica gel for on-field detection. The biosensor kit was stable for 20 days when stored at 4 °C. Moreover, a 1-(1-naphthylmethyl)-piperazine (NMP) efflux pump inhibitor can improve the sensitivity by 57 %.
- Published
- 2020
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