In order to enhance the application value of zein and improve the environmental pollution caused by traditional petroleum base. The composite antibacterial film was prepared by tape casting with zein, agar and carrageenan as film-forming substrate, gelatin as enhancer, glycerol plasticizer, natamycin and nisin as antibacterial agents, and its physicochemical properties and fresh-keeping effect were studied. The results show that, The Zein-AG film were in three composite antimicrobial film with mechanical strength of 7.12±0.91 MPa and light transmittance of 25.40±1.00%; The Zein-AG film has a high air permeability and water solubility; The Ze-CA/AG film showed strong antibacterial properties and showed significant differences compared with the other composite film (P <0.05), the antibacterial circle diameter of Aspergillus niger and Rhizobium is 35.155±0.85 mm and 23.17±0.94 mm, and it also can effectively inhibit the reproduction of S. aureus and E. coli.. In terms of preservation, Ze-CA / AG film has a good effect on the control of the weight loss rate, hardness and nutritional indicators of grapes, which is conducive to extending the preservation period of grapes. The research provides reference for the development and utilization of Zein and the preparation of green packaging materials.