Coaxial electrospinning technique was used to fabricate the core-sheath composite nanofibers of ZnO nanoparticle (Nps) (10%, 20% w/w) doped polymethyl methacrylate (PMMA) (as sheath) and polyvinyl alcohol (PVA) (as core). Fourier transform infrared (FT-IR) spectra were confirmed the weak forces arise between ZnO Nps, PMMA and PVA matrixes. The hexagonal (wurtzite) structure of ZnO Nps with similar to 30.8nm of diameter was confirmed from the X-ray diffraction pattern. The morphology and microstructure of core-sheath composite nanofibers were confirmed from the scanning electron microscopy (SEM) and transmission electron microscopy (TEM). It is clearly seen from the TEM images that the PMMA encapsulate the PVA core. Core-sheath composite nanofibers were assessed against Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis) bacteria through quantitative, disk diffusion and viable cell count methods. It was found that ZnO Nps doped core-sheath nanofibers were effectively inhibit the growth of gram positive bacteria, B. subtilis.