Abstract The present study was aimed towards green synthesis of gold nanoparticles (AuNPs) using fungal strain (SGSK-7) with assessment of their size using different techniques viz., X-ray diffractormeter, Dynamic light scattering, Atomic force microscopy, FE-Scanning Electron Microscopy and Fourier transform Infrared Spectroscopy. The intense characteristic diffraction peaks at angle 2Ѳ (38.13˚) presented the crystalline nature of the reduced gold solution and calculation of crystalline size (12 nm) using Scherer’s equation further confirmed synthesis of gold nanoparticles while dynamic light scattering analysis indicated the particle size range of gold nanoparticles between 2 nm to 50 nm. Field Emission Scanning Electron Microscopic (FESEM) and Atomic Force Microscopy (AFM) analysis of extracellular gold nanoparticles depicts spherical shape of AuNPs of approximately 40 nm. Fourier transform Infrared (FTIR) Spectroscopy depicted the presence of the gold nanoparticles along with the other biomolecules secreted by fungus which plays vital role in reduction, capping and stabilization of gold nanoparticles. Energy dispersive X-ray, mapping analysis, Zeta potential and UV-Vis spectrophotometery analysis further confirmed the presence and stability of gold nanoparticles, respectively. Zeta potential of gold nanoparticles leads to conclude the neutral nature (-10 mV to 10mV) of the particles. The UV-Visible spectrophotometeric analysis also confirmed the formation of gold nanoparticles with an absorption peak at 547 nm. On the basis of UV-Visible spectra the optical band gap of 2.44 eV is examined. The phylogenetic relatedness of SGSK-7 revealed 99% homology with Aspergillus tamari after sequencing of the ITS region followed by BLAST.