In the present study, we formulated and characterized CMC/PVA/CuO bionanocomposites to evaluate their use in coating processed cheese. Copper oxide nanoparticles (CuO-NPs) were prepared and added to a mixed solution of carboxymethyl cellulose (CMC)/polyvinyl alcohol (PVA) using compositions of 0.3, 0.6 and 0.9% (w/v). The CMC/PVA/CuO bionanocomposites were prepared by a solution casting method and used for coating processed cheese. The fabricated bionanocomposite films and CuO-NPs were characterized by TEM, SEM, EDEX, XRD, DLS, and FT-IR analysis. Inclusion of CuO-NPs decreased the gas transmission rate (GTR) and water vapor transmission rate (WVTR) of the prepared film. Also, the bionanocomposite suspensions exhibited high but variable inhibitory effects against several pathogenic bacteria and fungi. The impact of coating of processed cheese surfaces with the prepared bionanocomposite films on microbiological, physicochemical, textural and sensory properties of the processed cheese were assessed during 6 months of cold storage. Coating cheese with film containing CuO-NPs eliminated mould growth on the cheese surface and decreased significantly (P < 0.05) the total bacterial count of the cheese. Furthermore, coating of cheese decreased the moisture losses and retarded the increase in the cheese hardness during storage. The highest acceptability at the end of the storage period was given for processed cheese coated with the bionanocomposite containing 0.9% CuO-NPs. Thus, the obtained CMC/PVA/CuO bionanocomposite films could be a promising candidate for cheese packaging applications.