Novel nanocomposite based on Polyvinyl alcohol (PVA), Carboxymethyl Cellulose (CMC) and Zinc Oxide Nanoparticles (ZnO-NPs), namely CMC/PVA/ZnO nanocomposites, was fabricated via solution casting methodology. ZnO-NPs were firstly prepared by hydrothermal method then different ratios of both CMC and PVA (50:50 and 60:40 v/v) were prepared to create novel blend, then ZnO-NPs was loadings with different concentrations to the prepared blend. The prepared ZnO-NPs were tested using X-Ray Diffraction pattern (XRD) and Transmission Electron Microscope (TEM). As well as CMC/PVA/ZnO nanocomposites were studied using Fourier Transform Infrared (FT-IR), TEM, XRD, UV/Vis spectroscopy and TGA. The films containing CMC/PVA in a ratio of (50:50 v/v) displays best morphological, thermal, mechanical antibacterial properties than films prepared using (60:40 v/v). The prepared nanocomposites displayed respectable antibacterial activity against gram positive (Staphylococcus aureus), gram negative (Pseudomonas aeruginosa, Escherichia coli) bacteria and fungi (Candida albicans). Furthermore, the fabricated nanocomposites can be used as good materials for packaging applications.