In the last two decades, nanomaterials have received much attention for the treatment of multidrug-resistant microbes that threaten human health. In the current study, the novelty and scientific significance concentrated on the biogenic synthesis of bimetallic silver-zinc oxide nanoparticles (Ag-ZnO NPs) using pomegranate peel extract (PPE) for the first time. The new constructed bimetallic Ag-ZnO NPs possessed the synergistic activity at a low concentration to avoid toxicity and elevate the superior potential. UV-Vis. characterization illustrated that Ag-ZnO NPs were small in size (15.8 nm), which was observed at 395.0 nm. The SEM image of Ag-ZnO NPs, incorporated with PPE, exhibited uniform Ag-ZnO NP surfaces with a clear surface appearance. It can be detected that Ag-ZnO NPs were isolated typically as a rounded particle across the PPE, which showed as brilliant NPs combined and stabilized with the prepared PPE. Results also revealed that Ag-ZnO NPs exhibited potential antibacterial activity toward Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, and Enterococcus faecalis where minimum inhibitory concentrations (MICs) were 62.5, 125, 15.62, 62.5, and 250 µg/ml. Likewise, Ag-ZnO NPs appeared antifungal activity against Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and Aspergillus brasiliensis, where MICs were 7.81, 31.25, 125, and 62.5 µg/ml, respectively. Moreover, Ag-ZnO NPs exhibited anticancer activities against MCF7 and Caco2, where IC₅₀ was 104.9 and 52.4 µg/ml, respectively. Additionally, these concentrations are safe in use where results of cytotoxicity on Vero normal cell line confirmed that IC₅₀ was 155.1 µg/ml. Overall, bimetallic Ag-ZnO NPs were for the first time, successfully biosynthesized using PPE; also, they had a promising antibacterial, antifungal, and anticancer activities.