In recent years, bio-metallic nanoparticles have been considered in medical science due to their biomedical properties. This research aimed to develop an eco-friendly, simple and facile process to synthesize green silver nanoparticles (AgNPs) using Trachyspermum ammi fruit extract. The synthesized silver nanoparticles were characterized using FT-IR, UV-Vis spectroscopy, XRD, DLS, and TEM. The minimum inhibitory concentration (MIC) values of synthesized AgNPs and T. ammi extract for five standard bacteria strains were determined by the broth microdilution method. The obtained AgNPs exhibited Surface Plasmon Resonance centered at about 415 nm with an average particle size calculated to be 50 nm. The mean particle size and surface charge of biosynthesized AgNPs using T. ammi extract investigated by DLS and zeta potential were 26.78 ± 1.24 nm and − 13.96 mV, respectively. Furthermore, green synthesized AgNPs show high and efficient antibacterial activity against E. coli ATCC 25922, P. aeruginosa ATCC 27853, K. pneumonia ATCC 9997, S. aureus ATCC 25923, and E. faecalis ATCC 29212 with MIC values of 19, 19.5, 75, 150, and 39 µg/ml, respectively. Noteworthy, the antibacterial results illustrated that green synthesized AgNPs possess significantly higher antibacterial potency than chemically produced silver nanoparticles. Our findings highlight the effective and efficient synthesis of silver nanoparticles by T. ammi fruit extract and its significant antibacterial activities.