Development and characterization of a polymer gel with an immobilized enzyme to measure L-glutamate.
Development and characterization of a polymer gel with an immobilized enzyme to measure L-glutamate.
复制标题
用于测量 L-谷氨酸的固定化酶聚合物凝胶的开发和表征。
DOI：
10.1016/0003-2697(87)90671-3
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发表时间：
1987
影响因子：
2.9
通讯作者：
Copenhagen,DR
中科院分区：
文献类型：
--
作者：
Korenbrot,JI;Perry,R;Copenhagen,DR
谷氨酸脱氢酶（GDH）用于酶电极中以测量l-谷氨酸。GDH共价固定在由聚丙烯酰胺和N-丙烯酰氧基琥珀酰亚胺共聚产生的亲水性、可渗透性和半刚性凝胶中。优化了以高效率和最小变性将GDH保留在凝胶中所需的实验条件。酶的辅助因子和辅酶，NADH，NAD，ATP，ADP，GTP，和ZnCl 2，在固定化过程中保护酶的能力进行了探索。在最佳的实验程序下，产生了功能行为可重复且持久的含酶凝胶。该凝胶对l-谷氨酸的反应速度快，延迟小于500 ms;特异性高，对l-谷氨酸的反应是对d-谷氨酸、d-或l-天冬氨酸和N-乙酰组氨酸的反应的1000倍;灵敏度高，可测量约3 μm的浓度。
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 μm can be measured.