饲用中性蛋白酶酶活的测定方法

The Determination Method of Neutral Protease Activity

本文介绍了福林酚法测定中性蛋白酶的原理、步骤，并重点研究了福林法测定中性蛋白酶在实际应用中的一些问题。研究表明，无论何种来源的固体蛋白酶都可以用0．3mol／L氯化钠作为浸提剂，浸提时间为30min；酶促反应的时间为5min，显色反应为20min；酶液合适的稀释倍数为最后测定酶活时OD值不超过0．5；不同厂家生产的酪素底物对测定结果有影响，底物浓度应保持在1％；酶促反应的温度和pH分别为40℃和7．0。

This article introduced neutral protease determination method - Folin method, and studied some problems in practical application. It showed that, the best extraction solvent for solid neutral protease was 0.3 mol/L NaC1, extraction time was 30 min; the enzymatic reaction time was 5 min and coloring time was 20 rain; the dilution multiple was the best when the OD value was lower than 0.5 ; the suhstrate from different factory had effect on activity determination, and substrate concentration must reach 1% ; the temperature and pH for enzymatic reaction was 40℃ and7.0.