固定化谷氨酸脱羧酶柱式反应器测定L-谷氨酸  

An Enzyme Column Reactor of Immobilized L-Glutamic Decaboxylased for Determination of L-Glutamic Acid

作　　者：吴国琪[1] 凌达仁[1] 王忱[1] 王钒[1] 宋国强[1] 

机构地区：[1]江苏石油化工学院应用化学系,常州213016

出　　处：《分析化学》1999年第7期759-763,共5页Chinese Journal of Analytical Chemistry

基　　金：江苏省自然科学基金

摘　　要：用羧甲基化的N,N’-甲叉双丙烯酰胺交联的烯丙基葡聚糖(简称CM-CADB)凝胶树脂为新型载体,研究了谷氨酸脱羧酶(GDC)在CM-CADB上的固定化与环境的依赖关系.确定了酶固定化最佳条件,研究了固定化GDC的活性与底物浓度、pH、温度的依存性,动态响应特性,热稳定性和寿命,求算了米氏常数.将固定化GDC酶柱与进样系统具CO_2气敏电极的离子活度分析器-计算机数据采集系统匹配,构成酶反应谷氨酸检测装置,测定了固定化GDC酶柱的能斯特线性响应曲线,线性回归方程为y=43.3x+181.6,r为0.9936.The imobilization of L-glutamic decaiboxylase(GDC) on the carboxymethyl-copolyiner of allyl dextran and N, N'-methylene-bisacrylamide (CM-CADB) ion exchange resin and the correlation of immobilization with the surroundings were studied. The condition for the immobilization of CDC was optimized with studying the buffer solutions and the various parmeters: adsorption equilibrium time, amount of the enzyme, pH, temperature, ionic strength and ion exchange capacity of CM-CADB resin. The dependence of the activity of immobilized GDC on the substrate concentration, pH, temperature and the dynamic response to L-glutamic acid, the thermostability and the longevity were investigated. The Michaelis constant was calculated. The enzyme column of immobilized GDC matching with the flow-injection-ion analyzer-data collection make up the original form of the apparatus of biosensor mesuring L-ghitamic acid. The Nemst linear response curve has been measured. The linear equation is y = 43. 3x+ 181.6 with the correlation coefficient of 0.9936.

关 键 词：固定化酶 谷氨酸脱羧酶 谷氨酸 发酵液 测定 

分 类 号：TQ922.1[轻工技术与工程—发酵工程]