Effect of polygodial on the mitochondrial ATPase of Saccharomyces cerevisiae.

The fungicidal mechanism of a naturally occurring sesquiterpene dialdehyde, polygodial, was investigated in Saccharomyces cerevisiae. In an acidification assay, polygodial completely suppressed the glucose-induced decrease in external pH at 3.13 microgram/ml, the same as the fungicidal concentration. Acidification occurs primarily through the proton-pumping action of the plasma membrane ATPase, Pma1p. Surprisingly, this ATPase was not directly inhibited by polygodial. In contrast, the two other membrane-bound ATPases in yeast were found to be susceptible to the compound. The mitochondrial ATPase was inhibited by polygodial in a dose-dependent manner at concentrations similar to the fungicidal concentration, whereas the vacuolar ATPase was only slightly inhibited. Cytoplasmic petite mutants, which lack mitochondrial DNA and are respiration deficient, were significantly less susceptible to polygodial than the wild type, as was shown in time-kill curves. A pet9 mutant which lacks a functional ADP-ATP translocator and is therefore respiration dependent was rapidly inhibited by polygodial. The results of these susceptibility assays link enzyme inhibition to physiological effect. Previous studies have reported that plasma membrane disruption is the mechanism of polygodial-induced cell death; however, these results support a more complex picture of its effect. A major target of polygodial in yeast is mitochondrial ATP synthase. Reduction of the ATP supply leads to a suppression of Pma1 ATPase activity and impairs adaptive responses to other facets of polygodial's cellular inhibition.

In vitro antifungal susceptibilities of Candida albicans and other fungal pathogens to polygodial, a sesquiterpene dialdehyde.

In vitro antifungal activities of polygodial were investigated against several fungal pathogens. Polygodial showed strong antifungal activity, comparable to amphotericin B, against yeast-like fungi Candida albicans, C. utilis, C. krusei, Cryptococcus neoformans, Saccharomyces cerevisiae and also filamentous fungi including Trichophyton mentagrophytes, T. ruburum, and Penicillium marneffei. Other strains such as Aspergillus fumigatus, A. flavus, P. chrysogenum, C. lipolytica, and C. tropicalis showed moderate to low susceptibility to polygodial. The anti-fungal activity of polygodial was generally not reduced by several susceptibility-testing conditions such as medium, incubation temperature, inoculum size, and medium pH. However, polygodial's antifungal activity was strongly increased at acidic conditions. Unlike amphotericin B, polygodial did not show any hemolytic activity and also its antifungal activity was not diminished in the presence of ergosterol. Based on killing kinetics against growing and nongrowing C. albicans, polygodial showed strong and rapid fungicidal activity.

Indole and (E)-2-hexenal, phytochemical potentiators of polymyxins against Pseudomonas aeruginosa and Escherichia coli.

Combinations of polymyxins and phytochemicals were tested for antimicrobial activity against two gram-negative bacteria. Various degrees of potentiation were found against Pseudomonas aeruginosa and Escherichia coli with (E)-2-hexenal and indole. Three-compound combinations were found to further increase the activity of polymyxin B sulfate and colistin methanesulfonate against both bacteria. Combinations with colistin against P. aeruginosa resulted in the highest degree of potentiation, with a 512-fold increase in colistin antimicrobial activity. These results indicate the potential efficacy of phytochemical combinations with antibiotics to enhance total biological activity.