In this study, the addition of oregano oil chitosan nanoparticles (OEO-CSNPs) was conducted to enhance the comprehensive properties of gelatin films (GA), and the optimal addition ratio of nanoparticles was determined for its application in the preservation of mullet. Oregano oil chitosan nanoparticles were organically combined with gelatin at different concentrations (0%, 2%, 4%, 6% and 8%) to obtain oregano oil–chitosan nanoparticle–GA-based composite films (G/OEO-CSNPs), and thereafter G/OEO-CSNPs were characterized and investigated for their preservative effects on mullet. Subsequent analysis revealed that OEO-CSNPs were uniformly dispersed in the GA matrix, and that G/OEO-CSNPs had significantly improved mechanical ability, UV-visible light blocking performance and thermal stability. Furthermore, the nanoparticles exhibited excellent antioxidant and antibacterial properties, and they improved the films’ suitability as edible packaging. The attributes of the G/OEO-CSNPs were optimized, the films had the strongest radical scavenging and lowest water solubility, and electron microscopy also showed nanoparticle penetration into the polymer when the concentration of OEO-CSNPs was 6% (thickness = 0.092 ± 0.001, TS = 47.62 ± 0.37, E = 4.06 ± 0.17, water solubility = 48.00 ± 1.11). Furthermore, the GA-based composite film containing 6% OEO-CSNPs was able to inhibit microbial growth, slow fat decomposition and protein oxidation, reduce endogenous enzyme activity, and delay the spoilage of mullet during the refrigeration process, all of which indicate its excellent potential for meat preservation application.