β-半乳糖苷酶活性检测方法的研究

Study on the Detection Method of Galactosidases Activity

β-半乳糖苷酶被广泛用于医药、临床、化学等领域,其酶活性的测定缺少标准方法。采用ONPG比色测定法测酶活性,通过单因素试验研究了影响酶活性的缓冲液pH、底物溶液浓度、反应温度、酶与底物反应比、反应时间等因素,采用mintab15响应面分析法,确认显著影响因子。采用正交实验进一步优化检测条件,确定出最佳酶活性测定条件:反应试剂pH为6.3,反应时间15分钟,反应温度为45℃。该检测方法稳定、准确、高效,可作为β-半乳糖苷酶活性检测方法推广使用。

β-Galactosidases is widely used in medicine,clinical,chemistry and other fields.There is no standard method to determine its enzyme activity.In this paper,ONPG colorimetric method is used to determine its enzyme activity,the factors affecting enzyme activity,such as pH of buffer solution,concentration of substrate solution,reaction temperature,reaction ratio of enzyme to substrate and reaction time were studied by single factor experiment.The optimum conditions for enzyme activity determination were determined as follows:pH=6.3,reaction time was 15 minutes,reaction temperature was 45°C.The method is stable,accurate and efficient,and can be used as a Galactosidases assay.