Research was carried out at the Green Nanotechnology Laboratory, University of Agricultural Sciences, Dharwad, Karnataka, with a specific emphasis on the biosynthesis of zinc nanoparticles using Pseudomonas and Actinobacteria. Collection and screening of Pseudomonas and Actinobacterial isolates were done at Microbial Genetics Laboratory, Department of Agricultural Microbiology, UAS, Dharwad. The ZnNPswere biosynthesized and characterized through UV-Visible spectroscopy, Particle Size Analyzer (PSA), Scanning Electron Microscope (SEM), Energy-dispersive X-ray spectroscopy (EDS), X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). The ZnNPs synthesized through Pseudomonas(AUDP209) and Actinobacteria(AUDT636)isolates exhibited peak values at 365 and 374 nm, with average sizes of 48.7 nm and 54.9 nm, respectively.SEM images revealed the spherical shape of the biosynthesized ZnNPs, while EDX confirmed the zinc ions presence in the biosynthesized samples and the crystalline structure of the ZnNPswas confirmed by XRD.FTIR spectroscopy confirmed the organic compounds present in the microbial extracts responsible for the capping and stabilizing agents for biosynthesis of ZnNPs.