Ultrasonication-assisted solution casting was used to prepare polyvinyl alcohol (PVA)/sodium carboxymethyl cellulose (CMC)/nano-ZnO/multilayer graphene nanoplatelet (xGnP) composite films; the performances (mechanical properties, water vapor permeability (WVP), biodegradability and antibacterial activity) of these films were investigated as a function of the ZnO NPs:xGnP mass ratio and ultrasonication time. Intermolecular interactions among ZnO NPs, xGnP and the PVA/CMC matrix were shown to improve WVP, while X-ray diffraction and scanning electron microscopy analyses revealed that the internal reticular structure of ultrasound-treated PVA/CMC/ZnO NPs/xGnP composite films was in a fluffier state than that of the untreated composite films and the PVA/CMC film. The incorporation of ZnO NPs and xGnP into the composite film reduced its tensile strength and elongation at break, and increased antibacterial activity and biodegradability. In addition, we carried out the experiment of strawberry preservation and measured weight loss ratio, firmness, content of total soluble solids and titration acid. Finally, the composite film of 7:3 had the best preservation effect on strawberries. Thus, the obtained results paved the way to develop novel biodegradable composite films with antimicrobial activity for a wide range of applications.