The conventional method of synthesizing gold nanoparticles (Au NPs) by chemical methods tends to cause an environmental concern; therefore there is an increasing tendency to use safe and nontoxic methods. The aim of the present study was to evaluate the anti-dermatophytic potential of biologically synthesized Au NPs using the cell free supernatant of Streptomyces sp. VITDDK3. The synthesized Au NPs were characterized using UV-visible spectra at 550 nm. XRD pattern analysis revealed peaks at 38.10° (111), 44.28° (200), 64.42° (220) and 77.37° (311) corresponding to the diffraction facets of Au. SEM analysis showed that the Au NPs were cubical in shape with an average size of 90 nm. The synthesized Au NPs were tested by the well diffusion method for antifungal activity (zones of inhibition) against Microphyton gypseum (10±0.47 mm) and Trichophyton rubrum (13±1.03 mm). © 2012 Published by Elsevier B.V. All rights reserved.