Istituto di Biochimica delle Proteine, CNR, Naples, Italy.
Anal Chem. 2011 Mar 1;83(5):1530-6. doi: 10.1021/ac102025z. Epub 2011 Feb 2.
Pesticides are the plague of modern times, although much needed in agriculture, causing damage to the entire ecosystem, including humans. The high operative costs and the requirement of specialized personnel for pesticide detection, incentive to develop alternative solutions such as the set up of cheap, rapid, and simple to use biosensors. In this work, we evaluate the possibility to use the esterase 2 from Alicyclobacillus acidocaldarius as a biosensor for the detection of specific organophosphate pesticides. With the recent demonstration of the very high affinity of esterase 2 toward paraoxon, a more complete analysis on the detection methods in water as well as in purposely contaminated fruit juices was carried out. The inhibitory effects of a wide range of other pesticides on esterase 2 were investigated, showing a better selectivity with respect to nonspecific reaction of acethylcholinesterases, the main target of organophosphate pesticides. The applied methodology allowed one to detect 2.75 × 10(-3) ppm of neurotoxic agent, comparable to the efficiency of other acethylcholinesterase-based biosensors. Finally, a raw biosensor, based on EST2 immobilization on a nitrocellulose membrane, was devised and tested for paraoxon detection, showing longtime stability, reproducibility, and sensibility.
农药是现代社会的瘟疫，尽管在农业中非常需要，但却对整个生态系统造成了损害，包括人类。高运营成本和对农药检测的专业人员的需求，促使人们开发替代解决方案，例如建立廉价、快速且易于使用的生物传感器。在这项工作中，我们评估了使用来自嗜酸热脂环酸芽孢杆菌的酯酶 2 作为检测特定有机磷农药的生物传感器的可能性。最近已经证明了酯酶 2 对氧乐果具有非常高的亲和力，因此我们对水和故意污染的果汁中的检测方法进行了更全面的分析。我们还研究了广泛的其他农药对酯酶 2 的抑制作用，结果表明，酯酶 2 相对于有机磷农药的主要靶标——乙酰胆碱酯酶的非特异性反应具有更好的选择性。所应用的方法允许检测到 2.75×10^-3 ppm 的神经毒剂，与其他基于乙酰胆碱酯酶的生物传感器的效率相当。最后，我们设计并测试了基于 EST2 固定在硝酸纤维素膜上的原始生物传感器，用于检测氧乐果，结果表明其具有长时间稳定性、重现性和灵敏度。
