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利用正交试验法研究5个因素对测定星豹蛛(Pardosa astrigera)头胸部乙酰胆碱酯酶(AChE)活性的影响,以建立酶活测定的最佳反应体系。结果表明,各因素对AChE的比活力影响由大到小的顺序为反应体系的pH值、反应时间、酶质量浓度、反应温度和底物浓度。测定AChE活性的最佳反应条件是:酶质量浓度为12 g·L~(-1),底物浓度为0.6 mmol·L~(-1),反应体系pH值为7.0,反应温度为30℃,反应时间为5 min。此外,4种杀虫剂对星豹蛛头胸部AChE活性的影响试验结果表明,灭多威、辛硫磷、高效氯氰菊酯、毒死蜱对AChE的抑制中浓度(IC_(50))分别为7.76×10~(-5)mol·L~(-1)、1.76×10~(-4)mol·L~(-1)、4.12×10~(-4)mol·L~(-1)和4.94×10~(-4)mol·L,并且上述4种杀虫剂对AChE的抑制具有明显的剂量关系。
The effects of five factors on AChE activity in head and chest of Pardosa astrigera were studied by orthogonal test to establish the optimal reaction system for enzyme activity determination. The results showed that the order of the influence of various factors on the specific activity of AChE was the pH value of the reaction system, the reaction time, the enzyme concentration, the reaction temperature and the substrate concentration. The optimum reaction conditions for the determination of AChE activity were as follows: enzyme concentration 12 g · L -1, substrate concentration 0.6 mmol·L -1, reaction pH 7.0 and reaction temperature 30 ℃ , The reaction time is 5 min. In addition, the effect of four kinds of insecticides on AChE activity in the spider’s head and chest showed that the inhibitory concentrations (IC 50) of AChE of methomyl, phoxim, beta-cypermethrin and chlorpyrifos were 7.76 × 10 ~ (-5) mol·L -1, 1.76 × 10 -4 mol·L -1, 4.12 × 10 -4 mol·L -1 and 4.94 × 10 ~ (-4) mol·L, and the four kinds of insecticides had a significant dose-dependent inhibition of AChE.