乙酰胆碱酯酶的定点突变法设计及其在生物传感器领域的应用

Site-directed Acetylcholinesterase Mutants for Biosensoring

有机磷和氨基甲酸酯类杀虫剂抑制乙酰胆碱酯酶(AChE)的天然生物活性。近年来,AChE作为生物识别元素的生物传感器在杀虫剂残留量检测领域受到了广泛的关注。简要分析了AChE结构与功能关系,综述了近年来通过定点突变法设计AChE以增加对各种杀虫剂的检测灵敏度及其在生物传感器领域的应用现状,指出多种灵敏的AChE突变体同时被应用于多样品及多分析物的平行检测体系是该领域进一步的发展方向。

Acetylcholinesterase （ACHE） is the target site for the two groups of widely used pesticides： organophosphates and carbamates. In recent years, the use of AChE as biological recognition element in biosensor technology has gained more and more attention, in particular with respect to pesticides detection. The use of heterologous expression systems to produce various AChEs in high amount, as well as its successful purification, not only provided with large amounts of purified, functional and stable AChEs at low costs, but also encouraged genetic engineering work aimed to increase the sensitivity of the enzyme to various pesticides by site-directed mutagenesis based on the advanced studies on the structure-function relationship of ACHE. Several mutants were shown to improve significantly sensitivity to different pesticides. Thus it also makes it possible that hundreds of hypersensitive mutants were combined together for parallel analysis.