Now a days Nanobiotechnology is the promising area of research which controls the materials at nano level, having tremendous application in the area of medicine. Nanoparticles contain remarkable antimicrobial property to overcome the bacterial resistance developed by the pathogenic bacteria to varied amounts of antibiotic drugs. Silver nanoparticles have advantage over other metallic nanoparticles as it posses broad range of bactericidal property. Here in this paper we have reported the extracellular biosynthesis of silver nanoparticles from Penicillium italicum isolated from the soil collected from the campus of Sathyabama University Chennai Tamil Nadu India. After this nanoparticles were characterized by various techniques followed by antibacterial activity in combination with Sparfloxacin. Formation of dark brown in conical flask to cell free extract indicates the formation of silver nanoparticles. Synthesis by addition of 1Mm AgNO3.UV vis spectrophotometer showed the absorption peak at 418nm indicates the formation of nanoparticles. Atomic Force Microscopy (AFM) showed that nanoparticles are spherical and size is around 40nm.Nanoparticles showed good antibacterial alone and enhances the bactericidal activity of Sparfloxacin studied during the present study.