The GO–ZnO nanocomplexes that polycrystalline ZnO particles were loaded onto graphene oxide (GO) were prepared conveniently through the one-step reflux method and their antibacterial activities against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were also investigated. The results showed, a complete polycrystalline ZnO particle in the complexes had a quasi-spherical structure with less than 200 nm in diameter, and it was formed from approximately 15 nm crystal hexagonal wurtzite with distinct lattice planes. These polycrystalline ZnO particles possessed greater specific surface areas and well dispersed on the surface of GO. Their antibacterial properties indicated when 0.28 and 0.42 mg mL−1 GO–ZnO nanocomplexes were added to E. coli and S. aureus physiological solutions, their bactericidal percentages were > 99.9 % and approximately 90 % in 0.5 h and 4 h, respectively, which exhibited favorable antibacterial activities against the two germs. The antibacterial performance of the nanocomplexes was achieved through the synergistic action between GO and polycrystalline ZnO particles.