Proton pumping ATPase mediated fungicidal activity of two essential oil components

This work evaluates the antifungal activity of two essential oil components against 28 clinical isolates (17 sensitive, 11 resistant) and 3 standard laboratory strains of Candida. Growth of the organisms was significantly effected in both solid and liquid media at different test compound concentrations. The minimum inhibitory concentrations (MICs) of Isoeugenol (compound 1) against 31 strains of Candida ranged 100-250 μg/ml and those of o -methoxy cinnamaldehyde (compound 2) ranged 200-500 μg/ml, respectively. Insight studies to mechanism suggested that these compounds exert antifungal activity by targeting H(+)-ATPase located in the membranes of pathogenic Candida species. At their respective MIC(90) average inhibition of H(+)-efflux for standard, clinical and resistant isolates caused by compound 1 and compound 2 was 70%, 74%, 82% and 42%, 42% and 43%. Respective inhibition of H(+)-efflux by fluconazole (5 μg/ml) was 94%, 92% and 10%. Inhibition of H(+)-ATPase leads to intracellular acidification and cell death. SEM analysis of Candida cells showed cell membrane breakage and alterations in morphology. Haemolytic activity on human erythrocytes was studied to exclude the possibility of further associated cytotoxicity.