Copper nanoparticles (CuNPs) embedded in polyvinylpyrrolidone (PVP) and polyethylene oxide (PEO) fiber-matrices were prepared through centrifugal spinning of PVP/ethanol and PEO/aqueous solutions, respectively. The prime focus of the current study is to investigate the antibacterial activity of composite fibers against Escherichia coli (E. coli) and Bacillus cereus (B. cereus) bacteria. During the fiber formation, the centrifugal spinning parameters such as spinneret rotational speed, spinneret to collector distance, and relative humidity were carefully chosen to obtain long and continuous fibers. The structural and morphological analyses of both composite fibers were investigated using scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, and thermogravimetric analysis. In the antibacterial test, PVP/Cu and PEO/Cu composite fibrous membranes exhibited inhibition efficiency of 99.98% and 99.99% against E. coli and B. cereus bacteria, respectively. Basically, CuNPs were well embedded in the fibrous membrane at the nanoscale level, which facilitated the inhibition of bacterial functions through the inactivation of the chemical structure of the cells. Such an effective antibacterial agent obtained from forcespun composite fibers could be promising candidates for biomedical applications.