D201大孔离子交换树脂固定糖化酶的研究

Study on Immobilization of Glycosylase on D201 Macroporous Ion-exchange Resin

在单因素实验基础上,本文以D201大孔离子交换树脂为载体进行糖化酶的固定,以酶活回收率为评价指标,分析了固定化时间、料液比(酶与载体的比例)、给酶量等因素在糖化酶固定化中的影响。通过响应面法分析得到固定化最佳工艺为:固化时间27 min,料液比1:1,给酶量1800 U/g。此条件下的酶活回收率为60.66%,响应面优化结果验证良好。此外,通过稳定性检测发现:固定化糖化酶在常温下的半衰期为5天,低温4℃时保存较佳,操作稳定性较差。

On the basis of single factor test,D201 macroporous ion-exchange resin was used as carrier to immobilize glucoamylase,and the recovery rate of enzyme activity was used as evaluation index,the effects of immobilization time,ratio of material to liquid(the ratio of enzyme to carrier)and amount of enzyme on the immobilization of glucoamylase were analyzed.Response surface methodology(RSM)was used to determine the optimum conditions of immobilization.The optimum conditions were as follows:the time of immobilization was 27 min,the ratio of material to liquid was 1:1,and the enzyme dosage was 1800 U/g.Under these conditions,the recovery of enzyme activity was 60.66%,and the result of response surface optimization was good.In addition,the half-life of immobilized saccharifying enzyme was 5 days at room temperature,it was better preserved at 4℃,and the stability of operation was worse.