Owing to their unique physical, chemical, and potential antibacterial characteristics, zinc oxide nanoparticles (ZnO NPs) are highly valuable for medical applications. In this study, we aimed to determine the antimicrobial activity of ZnO NPs. ZnO NPs extracted from Lactobacillus showed antifungal activity. The synthesis of ZnO NPs was characterized using Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and UV-Vis diffuse reflectance spectroscopy (UV-Vis DRS). Using the well-diffusion method, the inhibition zones of ZnO NPs against Malassezia globosa and Candida albicans were approximately 28-29 mm in diameter, which compares favorably with those of fluconazole and ketoconazole. SEM analysis revealed a hexagonal morphology, with particle sizes ranging from 56.14-64.97 nm. UV-Vis analysis showed an absorption peak around 300 nm. This study revealed the dose-dependent antifungal effects of ZnO NPs and suggested that their interaction with fungal cells disrupts cell membrane integrity, potentially leading to cell death.