In this study, polyethylene terephthalate/polypropylene (PET/PP) films were treated via atmospheric pressure plasma, assembled with chitosan and various preservatives and applied for antimicrobial food packaging. Surface properties of these obtained films were studied by contact angle measurement, atomic force microscopy (ATM), X-ray photoelectron spectroscopy (XPS), Fourier transformed infrared spectroscopy (FT-IR) and dynamic laser scattering (DLS). The above results showed that the surface hydrophilicity and roughness of the films increased after the plasma treatment. Besides, chitosan and the preservatives were successfully assembled onto the surface of the films. In addition, the antimicrobial activities of the films against three kinds of microorganisms (Staphylococcus aureus, Bacillus subtilis and Escherichia coli) were investigated and the results indicated that the inhibition ratios against B. subtilis and E. coli reached almost 100% while the inhibition ratios against S. aureus were lower than 85%. Moreover, the accumulative release profiles of the antimicrobial substances migrating from the assembled films into the release solutions revealed that their release speed increased with the increment of temperature and acidity, but decreased with enhancing the ionic strength regulated by sodium chloride or with lowering the ionic mobility regulated by sucrose.