Synthesis of zinc oxide nanoparticles on their antimicrobial activity utilizing banana peelings could be a potential candidate to develop nano-formulated natural products as an alternative, efficient, inexpensive, and environmentally safe method with specified properties. The study aimed to investigate the biological synthesis, characterization, and antimicrobial study of zinc oxide nanoparticles (ZnONps) against clinical pathogens Pseudomonas aeruginosa and Staphylococcus aureus using Musa acuminata × balbisiana peelings. Data revealed that varying concentration, temperature, and pH level can significantly influence the antimicrobial activity and plays crucial steps in the preparation of ZnONps. Research on nanoparticles as antibacterial agents can have an essential application in the food industry as antibacterial agents in food packaging and towards foodborne pathogens. Since this study deals with the preliminary investigation on banana peelings locally grown in the Philippines as capping and reducing agent through green chemistry, it can promote potential application in the Philippine food industry. By proper incorporation of nanoparticles into packaging materials it may inhibit or cause foodborne pathogens bacterial death.