Enzymatic microtiter plate-based nitrate detection in environmental and medical analysis

被引：33

作者：

Borcherding, H ^{[1
]}

Leikefeld, S ^{[1
]}

Frey, C ^{[1
]}

Diekmann, S ^{[1
]}

Steinrücke, P ^{[1
]}

机构：

[1] Inst Mol Biotechnol, D-07745 Jena, Germany

来源：

ANALYTICAL BIOCHEMISTRY | 2000年 / 282卷 / 01期

关键词：

nitrate reductase; nitrate; NO detection; Griess reaction; microtiter plate; Pseudomonas stutzeri;

D O I：

10.1006/abio.2000.4585

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Our microtiter plate assay is based on the enzymatic reduction of nitrate by dissimilatory nitrate reductase from Pseudomonas stutzeri [EC 1.7.99.4]. Exogenous redox mediators like methyl viologen, methylene blue, and cibachron blue were applied to reduce nitrate reductase. Concentrations of 0.02-0.9 mM nitrate can be detected with +/-6% standard deviation, by using a photometric Griess reaction for the formed nitrite. Nitrate reductase is stable in the pH range 6.5-9.0 and works in the temperature range 4-76 degrees C. The assay shows no interferences with salt content up to 1 M chloride or 11 mM chlorate, and serum albumin content up to 50 mg/ml, The time demand of our two-step procedure is 20 min/100 samples. Nitrate reductase could be conserved on site of the wells of microtiter plates for at least 6 months at room temperature, The nitrate assay was applied in environmental and consumer goods analysis, and for medical diagnostics in human plasma samples. (C) 2000 Academic Press.

引用

页码：1 / 9

页数：9

共 36 条

[1] RESPIRATORY NITRATE REDUCTASE FROM DENITRIFYING PSEUDOMONAS-STUTZERI, PURIFICATION, PROPERTIES AND TARGET OF PROTEOLYSIS [J].