Mechanisms of Antibacterial Action of Three Monoterpenes

TLDR

The study evaluates the antimicrobial efficacy of linalyl acetate, (+)menthol, and thymol against *Staphylococcus aureus* and *Escherichia coli* and relates the results to their lipophilicity and water solubility. The authors assessed membrane disruption by measuring carboxyfluorescein leakage from unilamellar vesicles of diverse lipid compositions and monitored terpene interactions with multilamellar vesicles using differential scanning calorimetry. The results suggest that (+)menthol, thymol, and linalyl acetate exert antibacterial activity by perturbing microbial membrane lipids, increasing permeability and leakage, with effects modulated by drug lipophilicity, water solubility, membrane lipid composition, and surface charge, and that the compounds may also penetrate cells to target intracellular sites.

Abstract

In the present paper, we report the antimicrobial efficacy of three monoterpenes [linalyl acetate, (+)menthol, and thymol] against the gram-positive bacterium Staphylococcus aureus and the gram-negative bacterium Escherichia coli. For a better understanding of their mechanisms of action, the capability of these three monoterpenes to damage biomembranes was evaluated by monitoring the release, following exposure to the compounds under study, of the water-soluble fluorescent marker carboxyfluorescein from unilamellar vesicles with different lipidic compositions (phosphatidylcholine, phosphatidylcholine/phosphatidylserine [9:1], phosphatidylcholine/stearylamine [9:1], and phosphatidylglycerol/cardiolipin [9:1]). Furthermore, the interaction of the terpenes tested with dimyristoylphosphatidylcholine multilamellar vesicles as model membranes was monitored by means of differential scanning calorimetry. Finally, the results were related to the relative lipophilicity and water solubility of the compounds examined. Taken together, our findings lead us to speculate that the antimicrobial effect of (+)menthol, thymol, and linalyl acetate may result, at least partially, from a perturbation of the lipid fraction of microorganism plasma membrane, resulting in alterations of membrane permeability and in leakage of intracellular materials. Besides being related to physicochemical characteristics of the drugs (such as lipophilicity and water solubility), this effect seems to be dependent on lipid composition and net surface charge of microbial membranes. Furthermore, the drugs might cross the cell membranes, penetrating into the interior of the cell and interacting with intracellular sites critical for antibacterial activity.