Nanotechnology is a rising field of science and technology that deals with the particles having size in the range of 1 to 100 nm. Copper oxide nanoparticles has many properties like antifungal activity, antibacterial activity, optical properties, conductive properties, etc. Due to its demand of diversified use, copper oxide nanoparticles were fabricated using ecofriendly and non-toxic Annona muricata stem extract. The extract with copper sulphate pentahydrate showed gradual change in the colour of the extract from brown to green which indicates the CuO nanoparticles synthesis. The fabrication is followed by characterization of CuO nanoparticles using UV-vis spectroscope, FTIR, XRD and SEM. The characterization showed roughly spherical shaped nanoparticles in the range of 100nm with high crystalline monoclinic phase. FTIR absorption spectra conclude that the compounds attached with copper oxide nanoparticles could be polyphenols with aromatic ring. The CuO nanoparticles exhibited antibacterial activity; it showed the maximum activity against E.coli (18 mm). Keywords: Annona muricata, copper sulphate pentahydrate, FTIR, nanomaterials, SEM, XRD.