@article{mbs:/content/journal/micro/10.1099/mic.0.000979, author = "Chouichit, Pakwilai and Whangsuk, Wirongrong and Sallabhan, Ratiboot and Mongkolsuk, Skorn and Loprasert, Suvit", title = "A highly sensitive biosensor with a single-copy evolved sensing cassette for chlorpyrifos pesticide detection", journal= "Microbiology", year = "2020", volume = "166", number = "11", pages = "1019-1024", doi = "https://doi.org/10.1099/mic.0.000979", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.000979", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "directed evolution", keywords = "immobilization", keywords = "aldehyde tag", keywords = "insecticide", keywords = "error-prone PCR", keywords = "pump inhibitor", 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 %.", }