胰蛋白酶的固定化及其性质研究

Studies on Immobilization of Trypsin and Properties of Immobilized Trypsin

以自制的脱乙酰壳多糖作载体,戊二醛为交联剂,对胰蛋白酶的固定化条件及其固定化酶的性质进行了研究。考查了交联剂的用量、pH值、以及载体与酶的比例等因素对胰蛋白酶固定化的影响。在所选择的固定化条件下,固定化酶的活性回收可达50％以上。同时研究了固定化胰蛋白酶的一些性质;最适温度60℃,最适PH8.0,Km值比可溶性酶升高,热稳定性、pH贮存稳定性以及在乙醇水溶液中的稳定性明显高于可溶性胰蛋白酶。在柱式反应器内,以2％酪蛋白为底物对,操作半衰期为40天。

In this paper,the method for immobilization of trypsin and the properties of immobilized trypsin were repeated. We used deacetylchitin as support, and glutar-aldehyde as crosslinking agent to prepare immobilized trypsin. Effect of glutaraldeh-yde concentration, pH value and the ratio of trypsin to deacetylchitin used in binding the enzyme to support was studied. An activity retention value of 50% has been reached. The optimum pH value of immobilized trypsin was 8.0, the optimum temperature was 60℃. The Km value for the immobilized enzyme thus obtained higher than that of the soluble enzyme. Stability of immobilized enzyme to temperature, pH and storage in ethyl alcohol was obviously increased than soluble enzyme. The operation half-life time was 40 days.