离子交换树脂固定化α－淀粉酶的研究

Study on immobilization of α-amylase by ion exchange resin

选择12种吸附和离子交换树脂对α-淀粉酶进行固定化,筛选出固定效果较好的D380大孔弱碱性丙烯酸系阴离子交换树脂为载体,通过先吸附后交联的方法固定化。通过实验对加酶量、交联剂浓度、吸附时间、吸附pH等能够影响固定化酶活力回收率的因素的研究和优化,得出较优的固定化条件为:戊二醛浓度0.1%、处理时间45min、加酶量3mg/g(蛋白量/载体)、酶液pH5.8、25℃、固定化处理时间为6～10h,获得的固定化酶活力可达80U/g(载体),并进一步对固定化后的酶学性质作了初步研究。

Alpha-amylase was immobilized on 12 kinds of industrial absorption and ion exchange resins. Among them, D380 showed excellent result for a-amylase immobilization. Then a-amylase was immobilized employing the method of absorption and cross-linking. The effects of the amount of α-amylase, concentration of glutaraldehyde, cross-linking time, reaction time and pH on immobilization were analyzed. The results revealed that the optimum conditions for a-amylase immobilization were as follows, the glutaraldehyde concentration, 0.1%; the time pretreated D380 with glutaraldehyde, 45min; the amount of α-amylase used, 3mg protein/g carrier （pH5.8）; the temperature and time for immobilization, 25℃ and 6-10h , respectively. The activity of immobilized enzyme was 80U/g carrier. The properties of immobilized enzyme were then further investigated.