Endophytic fungus inhabits such as biotopes. Many endophytic fungi and their metabolites have been reported to have insecticidal and fungicidal activity. Biological approach using an endophytic fungus is a novel way towards the development of safe, environmental friendly, viable and green method for the synthesis of silver nanoparticles in the field of nanotechnology. Several species of endophytes are able to synthesize nanoparticles with potential antimicrobial activity. In the present work, a single step environmental friendly approach is employed to synthesize silver nanoparticles. We have investigated biosynthesis of silver nanoparticles using endophytic fungus Aspergillus fumigatus isolated from the leaves of Cannabis sativa and have achieved rapid formation of silver nanoparticles in a short duration.Furthermore, the antimicrobial potential of AgNPs was systematically evaluated. The synthesized AgNPs could efficiently inhibit various clinically isolated pathogens like E. coli, S. albus, Klebsiella pneumoniae and Enterococcus sp. The current research opens a new avenue for the green synthesis of nano-materials.