New materials with good antibacterial activity and less toxicity to other species have attracted numerous research interests. Modified Hummers method was used for preparing graphene oxide (GO). Zinc oxide/graphene oxide (ZnO/GO) nanocomposites were synthesized with three different ratios (0.5:1, 1:1, and 2:1) by solution precipitation method. The ZnO/GO nanocomposites were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, Raman spectroscopy, Brunauer-Emmett-Tellerspecific surface area, and transmission electron microscopy image. The characterization results showed that ZnO nanoparticles with a mean size of 12-18 nm were randomly decorated on the surfaces and edges of GO sheets. ZnO/GO 1:1 with a high specific surface area of 65 m 2 /g was obtained. The antibacterial activity of ZnO, GO, and ZnO/GO was tested against Gram negative bacteria escherichia coli (E. coli) and Gram positive bacteria staphylococcus aureus (S. aureus) using well diffusion method. The test results confirmed that antibacterial activity of ZnO/GO was higher than that of GO and ZnO. Additionally, the ZnO/GO with the ratio of 1:1 is the strongest activity and more active against S. aureus than against E. coli and minimal inhibitory concentration (MIC) value of ZnO/GO 1:1 is 80 µg/mL for S. aureus and 160 µg/mL for E. coli. This novel nanocomposite could be used as a potential material for antimicrobial application.