A colorimetric assay for detecting haloalkane dehalogenase activity

被引：65

作者：

Holloway, P ^{[1
]}

Trevors, JT ^{[1
]}

Lee, H ^{[1
]}

机构：

[1] Univ Guelph, Dept Environm Biol, Guelph, ON N1G 2W1, Canada

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 1998年 / 32卷 / 01期

基金：

加拿大自然科学与工程研究理事会;

关键词：

chloride ions; colorimetric; dechlorination; haloalkane dehalogenase; microplate-based; pH indicator dye;

D O I：

10.1016/S0167-7012(98)00008-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A pH indicator dye-based colorimetric assay was adapted to measure haloalkane dehalogenase activity quantitatively in cell extracts of bacteria using the microplate format. The assay is based on a decrease in the pH of a weakly-buffered medium when protons and chloride ions are released by the enzyme. Our assay is more convenient than other instrumental or chemical assay methods presently used to monitor dechlorination reactions. In addition, our assay is rapid, quantitative, can be automated to handle a large number of samples using microplates, and uses inexpensive and nontoxic reagents. The assay may be adapted to monitor dechlorinating reactions mediated by other dehalogenase enzymes, (C) 1998 Elsevier Science B.V.

引用

页码：31 / 36

页数：6

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