Evaluation of novel ß-ribosidase substrates for the differentiation of Gram-negative bacteria

Butterworth, Lynne, Perry, John, Burton, M., Davies, Gideon, Reed, Robert and Gould, Frances Kate (2003) Evaluation of novel ß-ribosidase substrates for the differentiation of Gram-negative bacteria. Journal of Applied Microbiology, 96 (1). pp. 170-176. ISSN 1364-5072

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Official URL: http://dx.doi.org/10.1046/j.1365-2672.2003.02130.x

Abstract

Aims:  To synthesize novel substrates for the detection of β-ribosidase and assess their potential for the differentiation of Gram-negative bacteria.

Methods and Results:  Two novel chromogenic substrates, 3′,4′-dihydroxyflavone-4′-β-D-ribofuranoside (DHF-riboside) and 5-bromo-4-chloro-3-indolyl-β-D-ribofuranoside (X-riboside) were evaluated along with a known fluorogenic substrate, 4-methylumbelliferyl-β-D-ribofuranoside (4MU-riboside). A total of 543 Gram-negative bacilli were cultured on media containing either DHF-riboside or X-riboside. Hydrolysis of DHF-riboside or X-riboside resulted in the formation of clearly distinguishable black or blue-green colonies, respectively. Hydrolysis of 4MU-riboside was evaluated in a liquid medium in microtiter trays and yielded blue fluorescence on hydrolysis which was measured using fluorimetry. β-Ribosidase activity was widespread with 75% of strains, including 85·6% of Enterobacteriaceae, showing activity with at least one substrate. Genera that demonstrated β-ribosidase activity included Aeromonas, Citrobacter, Enterobacter, Escherichia, Hafnia, Klebsiella, Morganella, Providencia, Pseudomonas, Salmonella and Shigella. In contrast, strains of Proteus spp., Acinetobacter spp., Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus generally failed to demonstrate β-ribosidase activity.

Conclusions:  The novel substrates DHF-riboside and X-riboside are effective for the detection of β-ribosidase in agar-based media and may be useful for the differentiation and identification of Gram-negative bacteria.

Significance and Impact of the Study:  This is the first report describing the application and utility of chromogenic substrates for β-ribosidase. These substrates could be applied in chromogenic media for differentiation of Gram-negative bacteria.

Item Type: Article
Additional Information: First ever report of enzyme substrates for the detection of ribosidase activity in bacteria and first description of the taxonomic utility of ribosidase activity for the differentiation of medically important bacteria. Substrates subsequently patented and commercialised.
Uncontrolled Keywords: Gram-negative bacteria
Subjects: B900 Others in Subjects allied to Medicine
C700 Molecular Biology, Biophysics and Biochemistry
F100 Chemistry
Department: Faculties > Health and Life Sciences > School of Life Sciences > Applied Sciences
Depositing User: EPrint Services
Date Deposited: 05 Jan 2009 14:50
Last Modified: 16 Aug 2017 20:22
URI: http://nrl.northumbria.ac.uk/id/eprint/1576

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