Prevalence of antibiotic-resistant bacteria demands alternatives to antibiotics. Copper-based nanoparticles with a high antibacterial property may be a solution to the problem. It is, therefore, important to understand the mode of antibacterial action of the nanoparticles (NPs). Despite reports on induction of reactive oxygen species (ROS) in bacteria by copper and copper-oxide nanoparticles and involvement of such ROS in cell killing, it is still unclear (a) if surface modification of the nanoparticles by media organics has any role on their antibacterial potency and (b) whether the bactericidal effects of these NPs are 'particle-specific' or 'ion-specific' in nature. We address these issues for cupric oxide nanoparticle (CuO-NP) in this study. Instead of nutrient medium, when E. coli bacterial cells were suspended in saline (0.9% NaCl), CuO-NP had a more anti-bacterial effect, with MBC (minimum bactericidal concentration) value of 6 µg/mL, than in nutrient medium with MBC value of 160 µg/mL. Moreover, the lysine-modified CuO-NP in saline had MBC at 130 µg/mL. Thus, unmodified CuO-NP was more efficient killer than modified one. Our finding further revealed that in saline;CuO-NP had 'particle-specific' antibacterial effect through generation of ROS and consequent oxidative damage by lipid peroxidation, protein oxidation and DNA degradation in cells.