To investigate the action mechanism of 1-benzyl-1,4-diazepane (1-BD) as efflux pump inhibitor (EPI) in mutants: Δ or overexpressing AcrAB and AcrEF efflux pumps. Effect of 1-BD on: antibiotic potentiation, by microdilution method; membrane functionality, by fluorimetric assays; ethidium bromide accumulation, by fluorometric real-time efflux assay; AcrB expression, by quantitative photoactivated localization microscopy. 1-BD decreases the minimal inhibitory concentration of levofloxacin and other antibiotics and increase ethidium bromide accumulation in overexpressing efflux pumps but not in the Δ strain. 1-BD increases membranes permeability, without sensibly affecting inner membrane polarity and decreases transcription. 1-BD acts as an EPI in with a mixed mechanism, different from that of major reference EPIs.
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