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This study aimed to synthesize packaging films using bioactive ingredients. The composite film was prepared by blending octenyl succinate cassava starch ester (OSCS) with chitosan (CS) nano-ZnO and then adding ε-polylysine (ε-PL). The study also explored the effect of different concentrations of ε-PL on OSCS/CS/ZnO films. Fourier infrared spectroscopyand fluorescence microscopy revealed that the composite film was formed by both hydrogen bonding and a Schiff base reaction. The diffraction peaks of the original materials in X-ray diffraction disappeared after film formation, indicating good miscibility between the materials. Scanning electron microscope showed that the density of its structure increased with increasing the ε-PL content. The thermogravimetric analysis showed that the addition of ε-PL improved the thermal stability of the composite film to some extent. When used in cherry preservation, the bio-based modified starch film effectively reduced cherry decay, stem dryness, and weight loss, maintained surface color, and increased the soluble solid content.