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D. Alvarez-Ramirez, F. Cerda-Ramirez, N. Balagurusamy, Y. Garza-Garcia, A. D. Ilyina
Effect of colicin E1 on atp release from E.
coli strains for its selective detection by bioluminsecence assay
Abstract
The purpose of this study
was to demonstrate that the bioluminescent assay could be optimized for
selective E. coli detection by employing colicin E1 for ATP
release. Colicin E1 is a
bacteriocin, which is an extracellular proteinaceous antimicrobial
substance that exerts a bactericidal activity on the same or closely related
species. Considering the sensitivity of luciferase to various inhibitors, it
was proved that the colicin E1 did not inhibit luciferase at different
concentrations. Further, the luciferase activity was not affected by vanadate
ions (using as ATP-ase inhibitor) at 0.05 mM concentration or less. By means of
spectrophotometric assay, it was demonstrated that colicin E1 caused
only inhibition of E.coli proliferation without an effect on Pseudomona
aeruginosa, Enterococcus faecalis and other bacterial strains. It
was observed that in the presence of colicin without vanadate, ATP was not
detected. ATP quantification was possible after 30 minutes of incubation and
only in the presence of both vanadate and bacteriocin. The ATP/CFU relations
obtained with colicin/vanadate demonstrated a good correlation with the values
detected using dimethyl sulfoxide (DMSO) for the release of ATP.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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