小牛肠碱性磷酸酶活性检测方法的建立

目的]建立适用于科研用小牛肠碱性磷酸酶活性的检测方法。方法]通过模拟科研中小牛肠碱性磷酸酶的使用条件及缓冲液，并研究不同底物浓度、反应时间、显色时间、缓冲液pH等对酶活性检测的影响。结果]使用不同缓冲液对酶活性进行检测，其测量值差异较大。若使用添加硼酸的铁氰化钾溶液，则可以实现颜色的稳定显示。最佳水浴时间为10min，最佳底物浓度为0．04mol／L。随着温度的增加，对低浓度酶活性的影响不大，但对高浓度酶活性的影响逐渐增大。当缓冲液pH为7．O～8．0时，能够较好维持酶活性的稳定性。缓冲液粒子强度对酶活性的影响不大。结论]成功建立了科研用小牛肠碱性磷酸酶的活性检测方法。

Establishment of Activity Detection Method of Calf Intestine Alkaline Phosphatase

Objective] The research aimed to establish the detection method of the activity of alkaline phosphatise for scientific research. Method] The using conditions and buffer of calf intestine alkaline phosphatise in the scientific research were simulated and the effects of different substrate concentration, reaction time, ＂coloration time, buffer pH and other factors on the detection of enzyme activity were studied. Result] Different kinds of buffer had greater effects on the activity of alkaline phosphatise. When potassium ferricyanide solution with adding boracic acid was used, the coloration was stable. The best time of water bath was 10 min and the best substrate concentration was 0.04 mol/L. As the temperature increased,its effects on the activity of alkaline phosphatise at lower concentrations was little,but its effects on the activity of alkaline phosphatise at higher concentrations were greater. When the pH of buffer was 7.0 - 8.0, the stability of the activity of alkaline phos- phatise could be maintained. The granule intensity of buffer had little effects on the activity of alkaline phosphatise. Conclusion] The detection method of the activity of calf intestine alkaline phosphatase for scientific research was successfully established.