The synthesis of metal and semiconductor nanoparticles is an expanding research area due to the potential applications in the development of novel technologies. In the present study, a simple and eco-friendly synthesis of zinc oxide nanoparticles (ZnO NPs) using leaf extract of Mentha pulegium L. has been used. The biosynthesized ZnO NPs were characterized various techniques such as UV-Vis absorption spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), Transmission electron microscopy (TEM), energy dispersive X-ray analysis (EDX) and Fourier transform infrared spectroscopy (FT-IR). The XRD data showed the crystalline nature of the nanoparticles and EDX measurements indicated the high zinc content of 56.26% and also oxygen with 43.74%. FT-IR confirmed the presence of functional groups of both leaf extract and ZnO NPs. The particles size and morphology determined from FE-SEM and TEM and UV visible absorbance spectrum of ZnO NPs exhibited the absorbance band at 370 nm. The synthesized ZnO nanoparticles as potential antibacterial agent has been studied on Escherichia coli and Staphylococcus aureus. These results indicate that aqueous extract of Mentha pulegium (L.) are effective reducing agents for green synthesis of ZnO NPs with significant antimicrobial potential.