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Detection of potential AcrAB-TolC multidrug efflux pump inhibitor in calyces extract of Hibiscus sabdariffa

Nehaya Al-Karablieh, Luay Abu-Qatouseh, Talal Aburjai.


Abstract

Aim: The aim of this study is to investigate occurrence of potential efflux pump inhibitor (EPI) against AcrAB-TolC efflux pump in the methanol extract of H. sabdariffa. Materials and Methods: Calyces of H. sabdariffa were purchased from the local market in April 2014, methanol extract of H. sabdariffa was subjected to agar plate diffusion against Escherichia coli TG1 and its ∆acrB-∆tolC and thin layer chromatography (TLC) bioassay. The corresponding EPI fraction was eluted by methanol. The synergistic effect of antimicrobials and EPI fraction was measured by minimum inhibitory concentration (MIC) determination for E. coli and Erwinia amylovora strains, and the ability of EPI fraction to enhance EtBr accumulation was conducted. Results: E. coli TG1 was more sensitive to the methanol extracts of H. sabdariffa than E. coli ∆acrB-∆tolC, and inhibition zone corresponding to flavones on TLC bioassay plate has been formed which might be related to the fraction of potential EPI. The MIC values revealed that EPI fraction enhanced the activity of the used antimicrobials by 4 to 8 folds in E. coli TG1 and by 4 to 10 folds in E. amylovora 1189. Addition of EPI fraction in a dose-dependent manner increased the intercellular accumulation of Ethidium Bromide (EtBr) in the wild type stains of E. coli TG1 and E. amylovora 1189. Conclusion: EPI fraction behaves like a multidrug efflux pump inhibitor and further investigation should be conducted for determination of the chemical structure of EPI fraction.

Key words: AcrAB-TolC, Efflux pump inhibitor, Hibiscus sabdariffa






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