In this study, copper oxide (CuO) nanoparticles were incorporate within polyvinyl alcohol (PVA) matrix and CuO–PVA nanocomposite film was fabricated by solution casting technique. In order to synthesize nanoparticles, a cost-competitive and facile approach namely mechanochemical method was implemented. As nanoparticles precursors, pre-determined amount of CuSO4, NaCl, and NaOH were mixed. The mixture was ball-milled in zirconia medium for 10 h at ambient temperature and rotational speed of 300 rpm. The XRD characterization analysis indicated CuO nanoparticles formation by ball milling. The CuO size was estimated to be in the ≈ 37 nm range. SEM images demonstrated the spherical morphology and little agglomeration for the nanoparticles. Furthermore, the bandgap energy of CuO was determined to be 1.84 eV. The CuO content in the nanocomposite was set to be 2, 4, 8, and 10 wt%. Strong interaction between the PVA matrix and CuO nanofiller was confirmed by XRD and FT-IR investigations. Moreover, integration of CuO to the polymer matrix resulted in a decreased PVA crystallinity. The antibacterial behavior of PVA was increased by the addition of nanoparticles, and the CuO–PVA nanocomposite containing up to 4 wt% CuO had desirable antibacterial properties demonstrated against Escherichia coli activity. CuO nanoparticles incorporating to PVA network led to reinforced mechanical and dielectric properties for the nanocomposites. Besides, nanoparticles loading had direct impact on the intensified properties. Tensile strength was enhanced from 85.8 (pure PVA) to 154.9 MPa (PVA-10 wt% CuO). The tensile modulus for neat PVA was 0.9 GPa and raised dramatically to 4.5 GPa for PVA-10 wt% CuO. Dielectric constant also improved more than three times at 1 kHz comparing PVA and PVA-10 wt% CuO.