Korenbrot J I, Perry R, Copenhagen D R
Anal Biochem. 1987 Feb 15;161(1):187-99. doi: 10.1016/0003-2697(87)90671-3.
Glutamate dehydrogenase (GDH) is used in an enzyme electrode to measure L-glutamate. GDH is covalently immobilized in a hydrophilic, permeable, and semirigid gel produced by the copolymerization of polyacrylamide and N-acryloxysuccinimide. Experimental conditions necessary to retain GDH in the gel with high efficiency and minimum denaturation are optimized. The abilities of enzymatic cofactors and coenzymes, NADH, NAD, ATP, ADP, GTP, and ZnCl2, to protect the enzyme during immobilization are explored. Under optimum experimental procedures an enzyme-containing gel is produced that is reproducible and long lasting in its functional behavior. The gel responds to the presence of L-glutamate with high velocity, the delay being less than 500 ms; high specificity, being 1000-fold more responsive to L-glutamate than D-glutamate, D- or L-aspartate, and N-acetylhistidine; and high sensitivity, a concentration of about 3 microM can be measured.
谷氨酸脱氢酶（GDH）用于酶电极中测量L-谷氨酸。GDH通过聚丙烯酰胺和N-丙烯酰氧基琥珀酰亚胺的共聚反应共价固定在一种亲水性、渗透性和半刚性的凝胶中。优化了以高效率和最小变性将GDH保留在凝胶中的必要实验条件。探索了酶辅因子和辅酶（NADH、NAD、ATP、ADP、GTP和ZnCl2）在固定化过程中保护酶的能力。在最佳实验步骤下，制备出一种含酶凝胶，其功能行为具有可重复性且持久。该凝胶对L-谷氨酸的存在反应迅速，延迟小于500毫秒；具有高特异性，对L-谷氨酸的反应性比对D-谷氨酸、D-或L-天冬氨酸以及N-乙酰组氨酸高1000倍；并且具有高灵敏度，能够测量约3 microM的浓度。
