We studied the structural and antimicrobial properties of copper oxide nanoparticles (CuO NPs) synthesized by a very simple precipitation technique. Copper (II) acetate was used as a precursor and sodium hydroxide as a reducing agent. X‐ray diffraction patter (XRD) pattern showed the crystalline nature of CuO NPs. Field emission scanning electron microscope (FESEM) and field emission transmission electron microscope (FETEM) demonstrated the morphology of CuO NPs. The average diameter of CuO NPs calculated by TEM and XRD was around 23 nm. Energy dispersive X‐ray spectroscopy (EDS) spectrum and XRD pattern suggested that prepared CuO NPs were highly pure. CuO NPs showed excellent antimicrobial activity against various bacterial strains (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Enterococcus faecalis, Shigella flexneri, Salmonella typhimurium, Proteus vulgaris, and Staphylococcus aureus). Moreover, E. coli and E. faecalis exhibited the highest sensitivity to CuO NPs while K. pneumonia was the least sensitive. Possible mechanisms of antimicrobial activity of CuO NPs should be further investigated.