Thymol inhibits ergosterol biosynthesis in Nakaseomyces glabratus, but differently from azole antifungals

Nakaseomyces glabratus is considered a high priority of attention according to WHO, and also is an important yeast species due to its high rate of intrinsic/acquired resistance against fluconazole. This study aimed at the possible mechanisms of action of thymol, as the promising new antifungal agent, in N. glabratus. Thirty previously identified N. glabratus isolates were selected for investigation of the thymol susceptibility pattern. The antifungal susceptibility test was performed according to the Clinical and Laboratory Standards protocol published as M27-A2 document. Likely changes in the expression pattern of genes involved in the ergosterol biosynthesis pathway were assessed by Real-time PCR assay. The ultrastructure characteristics of thymol-treated yeasts and also the possible interactive proteins, as targets for thymol binding, were performed by transmission electron microscopy (TEM) and reverse molecular docking, respectively. Minimum inhibitory concentrations ranged between 32-128 µg/mL which were statistically significant between the fluconazole-susceptible and fluconazole-resistant yeast group (P