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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


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.
Moscow University Chemistry Bulletin.
2006, Vol. 47, No. 1, P. 59

Copyright (C) Chemistry Dept., Moscow State University, 2002
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