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本文研究在-78℃下,~(60)Coγ辐射引发亲水性单体甲基丙烯酸β-羟乙酯(HEMA)聚合固定化葡萄糖氧化酶(GOD),实验结果证明,控制辐射总剂量不超过1×10~4Gy,对葡萄糖氧化酶的活力损伤极微。载体HEMA浓度取为50％,包埋量是1.0ml GOD/10ml缓冲溶液(pH为5.6,0.06 mol/L磷酸缓冲液),可以得到既有弹性,又具有保存活力高的葡萄糖氧化酶固定化圆珠。辐照聚合HEMA温度越低,保存活力愈高。反应底物pH试验判明,固定化葡萄糖氧化酶的最适pH值与自然葡萄糖氧化酶的最适pH值一样,不过固定化葡萄糖氧化酶比自然酶更广泛适应于pH值的变化。固定化葡萄糖氧化酶动力学实验结果:米氏常数为k’m=1.42×10~(-2)mol比自然酶km=1.0×10~(-2)mol略高,说明了反应底物的扩散速度受到了一定的限制。活化能测定是13.7kJ/mol。
In this paper, the immobilization of glucose oxidase (GOD) catalyzed by ~ (60) Coγ radiation-initiated hydrophilic monomer β-hydroxyethyl methacrylate (HEMA) was studied at -78 ℃. The experimental results show that the total radiation dose More than 1 × 10 ~ 4Gy, minimal damage to glucose oxidase activity. The carrier HEMA concentration of 50%, the volume of 1.0ml GOD / 10ml buffer solution (pH 5.6,0.06 mol / L phosphate buffer), can be both elastic and high vitality of glucose oxidase immobilization Ball. The lower the temperature of irradiation polymerization HEMA, the higher the viability of preservation. The pH of the reaction substrate shows that the optimum pH value of immobilized glucose oxidase is the same as that of natural glucose oxidase, but the immobilized glucose oxidase is more widely adapted to natural pH than natural enzyme. The kinetic results of immobilized glucose oxidase showed that the Michaelis constant km = 1.42 × 10 ~ (-2) mol was slightly higher than that of natural enzyme km = 1.0 × 10 ~ (-2) mol, which indicated that the reaction substrate The rate of proliferation has been somewhat limited. The activation energy was measured at 13.7 kJ / mol.