Colorimetric assay for Lindane dechlorination by bacteria

A colorimetric microtitre plate-based assay that detects haloalkane dehalogenase activity was modified to detect dechlorination of γ-hexachlorocyclohexane (Lindane). Dechlorination is indicated by the colour change of phenol red from red to yellow, in a weakly buffered solution, as the solution beco...

Full description

Saved in:
Bibliographic Details
Published in:Journal of microbiological methods 2001-11, Vol.47 (2), p.181-188
Main Authors: Phillips, Theresa M, Seech, Alan G, Lee, Hung, Trevors, Jack T
Format: Article
Language:English
Subjects:
Bacteria
Bacteria - metabolism
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biodegradation, Environmental
Biological and medical sciences
Chlorides - metabolism
Colorimetric
Colorimetry - methods
Dechlorination
Enzyme kinetics
Ethylene Dichlorides - metabolism
Fundamental and applied biological sciences. Psychology
g-Hexachlorocyclohexane
Insecticides - chemistry
Insecticides - metabolism
Isolate screening
Kinetics
Lindane
Lindane - chemistry
Lindane - metabolism
Microbiology
Oxidation-Reduction
phenol red
phenolsulfonphthalein
Pseudomonas - metabolism
Systematics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A colorimetric microtitre plate-based assay that detects haloalkane dehalogenase activity was modified to detect dechlorination of γ-hexachlorocyclohexane (Lindane). Dechlorination is indicated by the colour change of phenol red from red to yellow, in a weakly buffered solution, as the solution becomes acidic due to HCl formed during dechlorination. Enzyme activity can be monitored by reading the absorbance of each well at 540 nm. Positive controls for the assay were the known Lindane-degrading microorganisms, Rhodanobacter lindaniclasticus and Sphingomonas paucimobilis UT26. Dechlorination in a scaled-up version of the assay was confirmed by GC/ECD detection of known metabolites of the test microorganisms from which the enzyme extracts were prepared. The assay was used to measure the rate of dechlorination in cell-free extracts of R. lindaniclasticus. It was also used to screen the cell-free extracts of 24 bacterial isolates, from a Lindane-contaminated soil, for Lindane dechlorination activity. Although no isolates tested positive, the assay represents a new inexpensive and rapid screening tool for the detection of Lindane-degrading microorganisms.
ISSN:0167-7012
1872-8359
DOI:10.1016/S0167-7012(01)00299-8