壳聚糖微球固定化葡萄糖氧化酶的研究

以壳聚糖微球为载体,戊二醛为交联剂,固定葡萄糖氧化酶,对葡萄糖氧化酶的固定化条件及固定化酶的各种性质进行了研究,确定了酶固定的最佳条件为0.1g壳聚糖微球与5ml5%戊二醛交联,固定6mg葡萄糖氧化酶,在此条件下酶活力回收可达60%。固定化酶的最适温度为50℃,最适pH为6.0,通过Lineweaver-Burk作图,确定动力学参数Km值为18.3mmol/L,表观米氏常数较游离酶有所降低,固定化酶的热稳定性较游离酶明显提高,该固定化酶具有良好的操作及保存稳定性。

Glucose oxidase was immobilized on chitosan microspheres with glutaraldehyde by cross -linking reaction,the immobilization conditions and characterization of the immobilized enzyme were studied.The optimal conditions for immobilization were as follows:0.1g chitosan microsphere was treated with5ml 5%glutaraldehyde,then6mg glucose oxidase was immobilized on to the carrier.Optimal temperature and pH for the immobilized enzyme were50℃and6.0respectively.Kinetic parameter,K m was 18.3mmol/L by Lineweaver-Burk method.The temperature stability of the immobilized enzyme was better than free emzyme,and the immobilized enzyme also showed good operation and storage stability.