胰蛋白酶的固定化及其应用研究

通过γ-（2，3-环氧丙氧）丙基三甲氧基硅烷将胰蛋白酶固定在硅胶载体表面制备固定化胰蛋白酶. 实验测试了pH值，温度，时间等因素对固定胰蛋白酶的影响，利用红外光谱、热失重等方法对固定化胰蛋白酶进行了表征；通过高效液相色谱法考察了在弱碱性条件下固定化胰蛋白酶与16种天然小分子物质的结合情况. 结果表明在25 ℃，pH=8.0，反应18 h的条件下，对胰蛋白酶在硅胶载体上的固定量最大；且固定化胰蛋白酶与黄酮类成分、生物碱类成分、萜类成分有良好的结合，对酸类及嘌呤类小分子物质不结合. 

Studies on Immobilization of Trypsin and Application of the Immobilized Enzyme

Trypsin was immobilized on silica gel carrier by chemical reaction method with γ-(2,3-epoxypropoxy) propyltrimethoxysilicane as coupling agent. The immobilized trypsin was analyzed by FT-IR and thermogravimetry assays. The influences of pH, temperature and time on the chemical reaction were investigated in this study. The immobilized trypsin affinity chromatography was used to absorb 16 natural products under weakly alkaline condition. The results showed that the maximum immobilized quantity of trypsin on the carriers are obtained under the condition 25 ℃, pH=8.0 and 18 h, and flavonoids, alkaloids and terpenoids are absorbed well, however acid and purine components are not combined on the immobilized trypsin.