In the present study, silver nanoparticles (AgNPs) were synthesized from C. glutamicum and their antibacterial activity has been investigated. Dried biomass of C. glutamicum was challenged with aqueous Diamine silver ([Ag (NH3 )2 ] ) containing 1mM AgNo3 at different pHconditions.Transmission electron microscopy (TEM) and Scanning electron microscope (SEM) analysis of the AgNPs were carried out. X- ray diffraction (XRD) results have shown that these nanoparticles exhibit a face-centered cubic crystal structure. Energy-dispersive X-ray (EDX) spectroscopy analysis for the confirmation of elemental silver was carried out for the detection of elemental silver. Fourier transform infrared (FTIR) spectra analysis shows that some functional groups, such as the carboxyl and amide of the proteins, were involved in the processes of bioreduction. Antimicrobial activity of the synthesized AgNPs against multi drug resistant bacteria and fungi was assessed by well diffusion method. AgNPs from C. glutamicum exhibited a potential antibacterial activity against the bacterial strains. The antimicrobial activity of silver nanoparticles was extremely good towards bacterial strains than the fungal strains. Thus, this study proved that the C. glutamicum ATCC13032 can produce AgNPs and the nanoparticles can be used as an effective antimicrobial substance.