Biosensors based on highly sensitive acetylcholinesterases for enhanced carbamate insecticides detection |
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Authors: | Bogdan Bucur Didier Fournier Jean-Louis Marty |
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Institution: | a University of Perpignan, Centre of Phytopharmacy, BIOMEM, 52 Av. Paul Alduy, 66860 Perpignan Cedex, France b IPBS-CNRS, Route de Narbonne, 31077 Toulouse, France c University of Bucharest, Faculty of Chemistry, Sos. Panduri, No. 90-92, 70326 Bucharest, Romania |
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Abstract: | This paper presents the construction of amperometric biosensors for the highly sensitive detection of carbamate insecticides based on the inhibition of acetylcholinesterase (AChE). This enzyme was immobilised by entrapment in an optimised sol-gel matrix on TCNQ-modified screen-printed electrodes. The enzyme activity was estimated by measuring the thiocholine produced by the enzymatic hydrolysis of the acetylthiocholine using TCNQ as mediator. Wild and genetically engineered AChEs from Drosophila melanogaster (Dm) were chosen for their high sensitivity towards insecticides, which substantially improves the LOD compared with cholinesterases from other sources. The wild type and three mutant enzymes were tested against three carbamate insecticides: carbaryl, carbofuran and pirimicard. The best LOD were obtained with the Y370A mutant for carbaryl (1 × 10−8 M), the E69W mutant for pirimicarb (2 × 10−8 M) and the I161V mutant for carbofuran (8 × 10−10 M). The biosensors were applied to the analysis of two potable water samples. |
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Keywords: | Recombinant acetylcholinesterase Carbamate insecticide Sol-gel immobilisation Disposable biosensor Screen-printed electrode |
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