A highly sensitive biosensor with a single-copy evolved sensing cassette for chlorpyrifos pesticide detection
Pakwilai Chouichit, Wirongrong Whangsuk, Ratiboot Sallabhan, Skorn Mongkolsuk, Suvit Loprasert
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 %.