Due to unique properties of the nanoparticles (NPs) with biocompatibility, their application as drug in drug delievery and diagnostics, the recent scientific branch nanotechnology has emerged as hope in modern medicine. Zinc oxide nanoparticles (ZnO NPs) have gained tremendous interest due to their potential use as chemotherapeutic and antimicrobial agents. They are included in the category of "generally recognized as safe (GRAS) metal oxide". There is an urgent need for developing additional sources of ZnO NPs. Therefore, in the present study 30 cyanobacterial extracts were screened for ZnO NPs synthesis.. The color change of the reaction mixture from blue to pale white indicated the synthesis of ZnO NPs. It was further confirmed by UV-Visible spectroscopy that showed the absorption peak at 372 nm. The SEM analysis during screening revealed that Oscillatoria sp. synthesized smallest ZnO NPs (~40 nm) that were further optimized for their higher yield by altering reaction conditions (pH, temperature, reaction time, concentration of extract and metal precursor). Best conditions for ZnO NPs synthesis are (0.02 M zinc nitrate, 10 ml of extract volume, pH 8, at 80 °C for 3 h). The NPs were purified through calcination at 350°C and characterized by UV-Vis, FTIR, XRD, SEM-EDAX, TEM, Zeta potential and DLS analysis. The comparative analysis of purified biogenic ZnO NPs with commercial chemically synthesized ZnO NPs (CS), exhibited their superior nature as antioxidant and anti-bacterial agent against both gram-positive and gram-negative bacteria. Synergistic effects of biogenic ZnO NPs and streptomycin additionally favored for their future use as a potential biomedical agent.