The peel aqueous extract of Punica granatum was utilized to fabricate zinc oxide nanoparticles (ZnO-NPs) as a green approach. The synthesized NPs were characterized by UV-Vis spectroscopy, Fourier transform infrared (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy, which was attached to an energy dispersive X-ray (SEM-EDX). Spherical, well arranged, and crystallographic structures of ZnO-NPs were formed with sizes of 10–45 nm. The biological activities of ZnO-NPs, including antimicrobial and catalytic activity for methylene blue dye, were assessed. Data analysis showed that the antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria, as well as unicellular fungi, was observed to occur in a dose-dependent manner, displaying varied inhibition zones and low minimum inhibitory concentration (MIC) values in the ranges of 6.25–12.5 µg mL–1. The degradation efficacy of methylene blue (MB) using ZnO-NPs is dependent on nano-catalyst concentration, contact time, and incubation condition (UV-light emission). The maximum MB degradation percentages of 93.4 ± 0.2% was attained at 20 µg mL−1 after 210 min in presence of UV-light. Data analysis showed that there is no significant difference between the degradation percentages after 210, 1440, and 1800 min. Moreover, the nano-catalyst showed high stability and efficacy to degrade MB for five cycles with decreasing values of 4%. Overall, P. granatum-based ZnO-NPs are promising tools to inhibit the growth of pathogenic microbes and degradation of MB in the presence of UV-light emission.