DEVELOPMENT OF A SUBSTRATE RECYCLE AMPLIFICATION SYSTEM FOR L-GLUTAMIC ACID ASSAY

被引：5

作者：

LUONG, JHT

MULCHANDANI, A

MALE, KB

机构：

[1] National Research Council Canada, Biotechnology Research Institute, Montreal, Que.

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 1991年 / 13卷 / 02期

关键词：

AMPLIFICATION; GLUTAMIC ACID; GLUTAMATE OXIDASE; GLUTAMIC OXALOACETIC TRANSAMINASE; HYDROGEN PEROXIDE PRODUCTION; SUBSTRATE RECYCLING;

D O I：

10.1016/0141-0229(91)90165-7

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A sensitive enzymatic assay based on substrate recycling was developed for detection of L-glutamic acid. The procedure was carried out using glutamate oxidase for conversion of L-glutamic acid to alpha-ketoglutarate, ammonia, and hydrogen peroxide. Alpha-ketoglutarate was then recycled to L-glutamic acid by reacting with L-aspartic acid in the presence of glutamic oxaloacetic transaminase. The coupled reactions were monitored by measuring the accumulated hydrogen peroxide spectrophotometrically. The magnitude of amplification was dependent upon the concentrations of L-aspartic acid, glutamate oxidase, glutamic oxaloacetic transaminase, and the reaction time. Compared to the assay without amplification, a maximal 26-fold increase in sensitivity was obtained. When applied to measure glutamic acid levels in soya sauce, onion soup mix, and blood plasma, the results agreed well with those determined by Beckman high-performance analyzer.

引用

页码：116 / 122

页数：7

共 15 条

[11] CSF AMINO-ACIDS AND PLASMA-CSF AMINO-ACID RATIOS IN ADULTS [J].