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

共 17 条

[1]

Belkin Shimshon, 1992, Biodegradation, V3, P299, DOI 10.1007/BF00129090

[2] DETERMINATION OF TRACE AMOUNTS OF CHLORINE IN NAPHTHA [J].

BERGMANN, JG ;

SANIK, J .

ANALYTICAL CHEMISTRY, 1957, 29 (02) :241-243

[3] BACTERIAL-DEGRADATION OF DICHLOROMETHANE [J].

BRUNNER, W ;

STAUB, D ;

LEISINGER, T .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1980, 40 (05) :950-958

[4] BACTERIAL DEHALOGENASES - BIOCHEMISTRY, GENETICS, AND BIOTECHNOLOGICAL APPLICATIONS [J].

FETZNER, S ;

LINGENS, F .

MICROBIOLOGICAL REVIEWS, 1994, 58 (04) :641-685

[5] CRYSTAL-STRUCTURE OF HALOALKANE DEHALOGENASE - AN ENZYME TO DETOXIFY HALOGENATED ALKANES [J].

FRANKEN, SM ;

ROZEBOOM, HJ ;

KALK, KH ;

DIJKSTRA, BW .

EMBO JOURNAL, 1991, 10 (06) :1297-1302

[6] CLONING OF 1,2-DICHLOROETHANE DEGRADATION GENES OF XANTHOBACTER-AUTOTROPHICUS GJ10 AND EXPRESSION AND SEQUENCING OF THE DHLA GENE [J].

JANSSEN, DB ;

PRIES, F ;

VANDERPLOEG, J ;

KAZEMIER, B ;

TERPSTRA, P ;

WITHOLT, B .

JOURNAL OF BACTERIOLOGY, 1989, 171 (12) :6791-6799

[7]

Janssen Dick B., 1994, Biodegradation, V5, P249, DOI 10.1007/BF00696463

[8] PURIFICATION AND CHARACTERIZATION OF HYDROLYTIC HALOALKANE DEHALOGENASE FROM XANTHOBACTER-AUTOTROPHICUS GJ10 [J].

KEUNING, S ;

JANSSEN, DB ;

WITHOLT, B .

JOURNAL OF BACTERIOLOGY, 1985, 163 (02) :635-639

[9]

*OR RES INC, 1990, CHL EL MOD 96 17B CO

[10] ISOLATION AND CHARACTERIZATION OF FLAVOBACTERIUM STRAINS THAT DEGRADE PENTACHLOROPHENOL [J].

SABER, DL ;

CRAWFORD, RL .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1985, 50 (06) :1512-1518

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