固定化Neutrase中性蛋白酶的研究

以壳聚糖为载体、戊二醛为交联剂固定化Neutrase中性蛋白酶。通过单因素实验，分析了壳聚糖浓度、戊二醛浓度、交联时间对微球制备的影响及戊二醛加入量对酶固定的影响。由正交实验确定制备固定化酶的最佳工艺参数为：壳聚糖浓度为3％、戊二醛与葡胺糖残基摩尔比为1：2、制备微球交联时间为1h，微球与酶振荡吸附12h，再加入2．5％戊二醛交联，使戊二醛最终浓度达到0．9％，制备得固定化中性蛋白酶活力为112．69U／g。固定化蛋白酶的热稳定性和对酸碱的稳定性均较游离中性蛋白酶有所提高。

Studies on Immobilization of Neutrase

Synthetic method of immobilized Neutrase enzyme prepared by reacting glutaraldehyde with chitosan was studied. Thee ffectso fc hitosanc oncentration,a dditiono f glutaraldehyde andc rosslinking timeo n preparationo fb eadsa nd onc oncentration of glutaraldehyde immobilization were analysed. Through perpendicular experiment, the optimized conditions of immobiliza- tionw ered etermineda s follows: chitosan concentration3 %,m olr ationo f glutaraldehydea ndg lucosaminer esidue1 :2,c rosslinking time 1h for preparation of chitosan beads, and adsorption time 12h. The final concentration of 2.5% glutaraldehyde was 0.9%. The activity of the immobilized Neutrase prepared under the optimal conditions was 112.69U/g. The thermostability and pH stability of immobilized enzyme were higher than those of soluble enzyme.