Leaf extract of in vitro raised plants of Ceropegia thwaitesii was assessed for the green synthesis of silver nanoparticles (AgNPs). The biosynthesized AgNPs were authorized by UV–vis spectrophotometer with surface plasmon resonance at 430nm. The scanning electron microscope (SEM), dynamic light scattering (DLS) analysis confirmed the particle size 100nm, and X-ray diffraction (XRD) confirmed the crystalline character of AgNPs. Further, Fourier transform infrared (FTIR) authorized the presence of triterpenoids and methoxy groups played an important reduction role in the synthesis process. It shows the significant antibacterial efficacy against Salmonella typhi, Shigella flexneri, Klbsiella pneumonia, Eschericia coli and others species. From the results, it is suggested that green synthesized AgNPs could be used effectively in future biomedical engineering.