Recent advances in nanotechnology and the synthesis of nanoparticles through biosynthesis have increased the urge in scientists than for chemical or physical methods. The biosynthesis method is the most significant method than a conventional method because of its eco-friendly, low cost and rapid synthesizing process. The present study describes the antibacterial activity of silver nanoparticles (AgNPs) synthesized from leaf extracts of Dypsis lutescens. The synthesis of AgNPs was confirmed by colour change from light yellow to brown colour. Further, the morphology of the biosynthesized nanoparticles, average size and presence of functional groups were characterized by UV – Visible spectroscopy (UV-Vis), X-Ray diffraction (XRD) and Fourier transform infrared spectroscopy, respectively. The UV spectra results show a strong resonance center and surface of silver nanoparticles at 450 nm. XRD studies revealed that the synthesized AgNPs show crystalline in shape. The FT-IR spectrum described the biological molecules which stabilize and form the silver nanoparticles in the aqueous medium. The average AgNPs size was found to be 31 nm by using the Debye-Scherrer formula. The antimicrobial property of AgNPs was tested against Escherichia coli (MTCC 443) and Vibrio cholerae (MTCC 3906) pathogen, which showed maximum zones of inhibition of 22 mm at a concentration of 100 µL. Therefore, the biosynthesized AgNPs proved to have significant antibacterial activity.