The precipitation process was used to synthesize ZnO nanoparticles (ZnO NPs), which were functionalized with fifteen amino acids and three surfactants. X-ray diffraction (XRD), field emission scanning microscopy (FESEM) with energy-dispersive X-ray spectroscopy, Fourier transformed infrared spectroscopy (FTIR), Raman spectroscopy, and UV-Vis spectroscopy were used to evaluate the synthesized and functionalized ZnO NPs. The characterization of the produced ZnO using XRD and FESEM revealed the development of a nanoscale hexagonal crystal, and the spectroscopic methods validated the presence of IR functional groups, Raman phase mode, and UV optical absorbance. The ZnO samples that had been functionalized demonstrated deformation in a surface morphology while maintaining chemical stability. The functionalization procedure effectively increases the inhibitory efficacy of NPs against Aspergillus Niger (Accession No. MZ435922) and Aspergillus Fumigatus (Accession No. MZ435863), as demonstrated by the antifungal activity. The findings of current work provide a foundation for improving the biological activity of NPs against fungi by modifying them with active medium and boosting their anti-microorganism and antioxidant activity.