The phytosynthesis of n-type Cadmium Oxide Nanoparticles reduces the toxicity of the substance and makes it Eco-friendly. This Eco-friendly biosynthesis of CdO NPs was synthesized for the first time from the Queen of herbs, Ocimum Sanctum (holy basil).The biosynthesized Cadmium oxide was prepared using Ocimum leaf extract as a reductant and Cadmium Chloride and hydroxide as cadmium and oxide source materials by Co- Precipitation method. Thus obtained Cadmium Oxide Nanoparticles were characterized by different techniques such as X-ray diffraction (XRD), Fourier Transform infrared spectroscopy (FTIR), Scanning electron microscope (SEM),Energy dispersive X-ray spectroscopy(EDS) to study the structural and morphological properties. XRD pattern exhibited the formation of face centered cubic structure of CdO NPs with an average crystalline size of 11.5nm .The chemical bond formation of CdO NPs were confirmed by FTIR spectrum in the range of (400-4000cm-1). The SEM micrographs revealed the predominant formation of Cauliflower shape with a particle size in the range of 61-142nm. The high purity of the biosynthesized nanoparticles were confirmed by EDS analysis. Further it was tested against gram positive and gram negative bacterial strains and showed significant antibacterial activity. This biosynthetic research study opens an innovative window to progress our understanding of how CdO NPs shows resistance to different bacterial strains.