Biosensors based on highly sensitive acetylcholinesterases for enhanced carbamate insecticides detection

被引:97
作者
Bucur, B
Fournier, D
Danet, A
Marty, JL
机构
[1] Univ Perpignan, Ctr Phytopharm, BIOMEM, F-66860 Perpignan, France
[2] CNRS, IPBS, F-31077 Toulouse, France
[3] Univ Bucharest, Fac Chem, Bucharest 70326, Romania
关键词
recombinant acetylcholinesterase; carbamate insecticide; sol-gel immobilisation; disposable biosensor; screen-printed electrode;
D O I
10.1016/j.aca.2005.12.060
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 (Din) 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 (I x 10(-8) M), the E69W mutant for pirimicarb (2 x 10(-8) M) and the It 6 1 V mutant for carbofuran (8 x 10(-10) M). The biosensors were applied to the analysis of two potable water samples. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:115 / 121
页数:7
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