In the present investigation, green synthesis of zinc oxide nanoparticles were successfully synthesized by biological method using aqueous stem extract of Ruta graveolens act as reducing agent. Formation of ZnO nanoparticles were characterized by powder X-ray diffraction (PXRD), UV-visible spectroscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) techniques. Zinc oxide nanoparticles were subjected to biological properties such as antibacterial and antioxidant studies. The PXRD pattern reveals that ZnO sample belongs to hexagonal phase with Wurtzite structure. The UV-vis absorption spectrum shows an absorption band at 355 nm due to ZnO nanoparticles. SEM images show that the particles have spherical like structure with large surface area and the average crystallite sizes were found to be in the range *28 nm. These observations were confirmed by TEM analysis. The ZnO nanoparticles are found to inhibit the antioxidant activity of 1,1-diphenyl-2-picrylhydrazyl free radicals effectively. ZnO Nps exhibit significant bactericidal activity against Gram -ve bacterial strains such as Klebsiella aerogenes, Pseudomonas aeruginosa, Escherichia coli and Gram ?ve Staphylococcus aureus by agar well diffusion method.