采用单因素和Box-Behnken实验对LX-1000EP树脂固定化戊二酰-7-氨基头孢烷酸酰化酶的工艺条件进行优化.首先通过单因素设计研究载体用量、缓冲液pH值、时间、温度四个因素对固定化戊二酰-7-氨基头孢烷酸酰化酶活力的影响,在此基础上,利用响应面试验设计对LX-1000EP树脂固定化戊二酰-7-氨基头孢烷酸酰化酶的条件进行优化.载体用量、缓冲液pH值、时间、温度对固定化戊二酰-7-氨基头孢烷酸酰化酶都有影响,固定化最佳工艺条件为:载体用量3.3 g、pH值7.9、时间40 h、温度25℃,在此条件下固定化酶活力达到(69.5±0.8) U/g,酶活回收率达到42.41％.将响应面法应用于固定化戊二酰-7-氨基头孢烷酸酰化酶条件的优化是可行的.%The immobilization technology conditions of GL-7-ACA acylase with LX-1000EP resin were optimized by using single factor analytical methodology and Box-Behnken experimental design.Firstly,the effects of carrier dosage,buffer solution pH,time and temperature on the vitality of GL-7-ACA acylase were investigated by adopting single factor design,based on which,the immobilization conditions were optimized by response surface methodology.The results show that the four factors all had effects on immobilized GL-7-ACA acylase activity,and the optimal immobilization conditions are as follows:3.3 of the carrier dosage,pH 7.9,time 40 hours and temperature 25 ℃.Under these appropriate conditions above,the immobilized enzyme,s activity was 69.5 ± 0.8U/g,the activity recovery was 42.41％.So it is feasible to optimize the immobilization conditions of GL-7-ACA acylase with LX-1000EP by using response surface methodology.
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