In this work, using Macrotyloma Uniflorum leaves an affordable and eco-friendly, ZnO and Ag-ZnO metal oxide nanoparticles were reported. Ensuing ZnO and Ag-ZnO nanoparticles were characterized by UV–visible spectroscopy, FT-IR Spectroscopy, X-ray diffraction, Scanning Electron Microscopy with Energy Dispersive Spectroscopy, Transmission Electron Microscopy, and Dynamic Light Scattering. The green synthesized ZnO and Ag-ZnO nanoparticles comprise an average size of about 120.16 nm and 91.17 nm respectively. The minimum inhibitory concentrations (MIC) of these ZnO and Ag-ZnO nanoparticles and mixtures thereof, Ag-ZnO, were determined on B.subtilis, Streptococci and E.coli cultures. MIC and their antimicrobial activity were studied in vitro; both types of nanoparticles showed high antibacterial activity. Also, it has shown excellent results with MIC value of 62.5 µg/ml for antibacterial activity against ZnO and Ag-ZnO nanoparticles. The Ag-ZnO nanoparticles were shown better antimicrobial effect than the ZnO nanoparticles. So, we can strongly suggest these green synthesized nanoparticles as a potent agent for biological applications.