辣根过氧化物酶活性检测方法研究

Detection Method of Horseradish Peroxidase Activity Colorimetry

通过单因素实验和正交实验研究了辣根过氧化酶(HRP)活性测定条件:缓冲溶液种类、缓冲溶液离子浓度、pH值、温度对其影响。实验结果表明在25℃下,0.1mol/L,pH=7.0的磷酸盐缓冲溶液中反应,辣根过氧化物酶的酶活力达到最大值1.13×10~7U/g;确定辣根过氧化物酶活性的最佳检测方法:1mg HRP溶解于1m L 0.1mol/L磷酸盐缓冲溶液(pH=7.0),反应温度为25℃,检测波长为436nm。该方法简单可靠,操作简便,可有效应用于实验室和常规生化检测。

The enzyme activity of HRP was studied by single factor experiment and orthogonal experiment.The enzyme activity achieves the maximum when the temperature is 25 ℃.In the orthogonal experiments,the results indicate that under 25 ℃,0.1 mol/L,phosphate buffer solution of pH7.0,HRP enzyme activity come to the maximum 1.13 × 10 7U/g.According to the experiments above,the best detection method of HRP activity comes out,which is in the case that 1 mg HRP dissolved 1 ml 0.1 mol/L phosphate buffer solution （pH7.0）,reaction temperature being 25 ℃,detection wavelength of 436 nm.The method is simple,convenience and remarkable,which can be applied to labor tests and routine biochemical tests.