In this study, we tried to enlighten the structure of zinc oxide nanoparticles (ZnO NPs) obtained from Thymbra Spicata L. plant by using green synthesis method in various ways. Some properties of zinc oxide nanoparticles were determined by using the characterization methods that scanning electron microscopy (SEM), Energy Dispersive X-ray analysis (EDX), fouirer transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), ultraviolet visible spectroscopy (UV-Vis) spectroscopy methods. The detected Zn nanoparticle sizes were determined to be between 6.5 nm and 7.5 nm. In addition to these studies, we investigated the antimicrobial effects of zinc oxide nanoparticles obtained by green synthesis against some pathogens. According to the results, it was seen that zinc oxide nanoparticles formed zones with a diameter of 16.3 mm, 10.25 mm, 13 mm and 10.2 mm, respectively, against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 25952, Pseudomonas aeruginosa ATCC 27853 bacteria and Candida albicans ATTC 90028 fungus, respectively. However, the radical quenching activity (DPPH) of the nanoparticles (Ts-ZnONP (79.67%)) was determined to be quite good compared to the positive control BHA. In addition, it is seen that the protective effect of ZnO NPs against DNA damage increases depending on the concentration. At a concentration of 100 mg/L, the DNA damage inhibitory effect was found to be maximum. In line with the comprehensive results, it was determined that the zinc oxide nanoparticles obtained with the green synthesis method have the potential of use in a wide variety of fields.