Synthesis and evaluation of novel flourogenic substrates for the detection of bacterial ß-galactosidase.

Chilvers, Kay Frances, James, Arthur, Perry, John and Reed, Robert (2002) Synthesis and evaluation of novel flourogenic substrates for the detection of bacterial ß-galactosidase. Journal of Applied Microbiology, 91 (6). pp. 118-1130. ISSN 1364-5072

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Abstract

Aims: A widely used coumarin derivative is 7-hydroxy-4-methylcoumarin-?-Dgalactoside (4-methylumbelliferone-?-D-galactoside; 4-MU-GAL). This galactoside is utilized as a substrate for the detection of the ?-galactosidase activity of coliform bacteria in water analysis. The intense fluorescence of coumarin-based molecules has enabled them to be incorporated into enzymebased tests for the quantitative assay of indicator bacteria. The aim of this present study was to evaluate the potential of other coumarin derivatives, by synthesis of a selection of core coumarin molecules. Methods and Results: Several coumarin derivatives were found to be more promising than 4-MU, with ethyl-7-hydroxycoumarin-3-caboxylate (EHC) giving a combination of greater fluorescence over a broad pH range and reduced growth inhibition with 12 representative coliform strains. On conversion to a ?-galactoside derivative, EHC-GAL generated a more rapid fluorescence than any other tested substrate. Conclusions: When tested in a broth assay format, based on most probable number (MPN), low numbers of coliforms were detected with EHC-GAL around 1 h earlier than with 4-MU-GAL. Significance and Impact of the Study: The present study suggests that EHC-GAL should be evaluated as a substrate for the detection of coliforms in water analysis, due to a combination of the following favourable features: (i) reduced toxicity; (ii) increased fluorescence; (iii) pH stability of fluorescence; and (iv) rapid detection.

Item Type:	Article
Additional Information:	This paper describes the synthesis of substrates designed to detect clinically important microorganisms. Prof. James designed and synthesised the substrates. The paper involved collaboration with BioMerieux.
Uncontrolled Keywords:	Microorganisms-Detection
Subjects:	C500 Microbiology
Department:	Faculties > Health and Life Sciences > Applied Sciences
Depositing User:	EPrint Services
Date Deposited:	23 Dec 2008 09:54
Last Modified:	31 Jul 2021 08:38
URI:	http://nrl.northumbria.ac.uk/id/eprint/87

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