Incorporating two different nanoparticles in nanocomposite films is promising as their synergistic effects could significantly enhance polymer performance. Our previous work conferred the remarkable antimicrobial (AM) properties of the polylactic acid (PLA)-based film using optimal formulations of synergistic graphene oxide (GO)/zinc oxide (ZnO) nanocomposites. This study further explores the release profile of GO/ZnO nanocomposite and their impact on the antimicrobial properties. A fixed 1.11 wt% GO and different ZnO concentrations were well dispersed in the PLA matrix. Increasing ZnO concentrations tended to increase agglomeration, as evident in rougher surfaces. Agglomeration inhibited water penetration, leading to a significant reduction in water permeability (46.3 %), moisture content (31.6 %) but an improvement in Young's Modulus (52.6 %). The overall and specific migration of GO/ZnO nanocomposites was found to be within acceptable limits. It is inferred that the release of Zn2+ ions followed pseudo-Fickian behavior with an initial burst effect. AM film with the highest concentration of ZnO (1.25 wt%) exhibited the highest inhibition rate against Escherichia coli (68.0 %), Bacillus cereus (66.5 %), Saccharomyces cerevisiae (70.9 %). Results suggest that GO/ZnO nanocomposites with optimal ZnO concentrations have the potential to serve as promising antimicrobial food packaging materials, offering enhanced barrier, antimicrobial properties and a controlled release system.