葡萄糖氧化酶-过氧化氢酶共固定研究

以弱碱性阴离子交换树脂D301T为载体，对过氧化氢酶（CAT）和葡萄糖氧化酶（GOD）两种酶进行分次固定，并对固定化条件进行了优化。所得最佳工艺条件为：共固定化酶采用先固定CAT，再固定GOD的顺序进行，其中CAT 0.5 mL，GOD 0.25 mL。所得CAT蛋白结合量为1.07 mg/g，固定化效率为46.71%；GOD蛋白结合量为1.58 mg/g，固定化效率为43.62%；每mL GOD酶液的表观酶活为47.98 U/mL，每g载体中GOD的酶活为12.0 U/g。以戊二醛作为交联剂，体积分数取0.5%，交联时间取15 min时，所得固定化酶表观酶活达到最大值，为14.66 U/g，固定化酶连续反应10批后，其酶活为初始值的85.3%，显示出固定化酶具有良好的操作稳定性。

Study of the co-immobilization of glucose oxidase and catalase

Glucose oxidase(GOD) and catalase(CAT) have been sequentially co-immobilized with D301T resin as the carrier.The effects of varying the immobilization sequence and the initial amounts of GOD and CAT on the GOD activity were determined.The results showed that the highest activity was observed when CAT was first immobilized on the carrier followed by immobilization of GOD,with initial amounts of 0.5 mL and 0.25 mL,respectively.The maximum protein loading of CAT was 1.07 mg/g and that of GOD was 1.58 mg/g.The co-immobilization degree of CAT was 46.71% and that of GOD was 43.62%.The apparent enzyme activity of the product was 47.98U/mL liquid enzyme and 12.0U/g carrier.The highest apparent enzyme activity was 14.66U/g carrier when glutaraldehyde was used to cross-link the enzyme and carrier.The concentration of glutaraldehyde used in this paper was 0.5% and the reaction time was 15 min.After reusing 10 times,the residual activity was about 85.3% of the initial activity,which showed the excellent operational stability of immobilized GOD-CAT.