星豹蛛乙酰胆碱酯酶活性测定条件的优化

Study on the Optimal Conditions for Analyzing Activity of Acetylcholinesterase Derived from Pardosa astrigera

为了进一步研究星豹蛛乙酰胆碱酯酶的生物活性,利用正交试验研究了反应体系的pH值、反应时间、酶浓度、反应温度和底物浓度5个因素对测定星豹蛛(Pardosa astrigera L.Koch)头胸部乙酰胆碱酯酶活性的影响,以优化酶活性测定的最适条件。对试验结果进行极差和方差分析,确定了星豹蛛头胸部乙酰胆碱酯酶的最适反应条件是:酶浓度为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。

To study the resistance status of Pardosa astrigera and its acetylcholinesterase（AChE） characeristics systemically,to establish a quick,accurate method,we used the orthogonal experiment and took five factors for the experiment,to assay the activity of acetylcholinesterase from the cephalothorax of Pardosa astrigena.The concentrations of acetylcholinesterase and substrate,pH and temperature of the reaction system,and reaction time were optimized.As a result,it was found that the effects on the activity of acetylcholinesterase were in the order of pHtimeAChE temperatureconcentration of substrate.The optimal conditions were： 12 g·L^-1,0.6 mmol·L^-1 substrate,pH 7.0,30 °C,5 minutes.