The development of methodologies for the synthesis of nanoparticles of well-defined size and shape is a challenging one and constitutes an important area of research in nanotechnology. This Full Paper describes the controlled synthesis of multishaped gold nanoparticles at room temperature utilizing a simple, green chemical method by the interaction of chloroauric acid (HAuCl4 · 3H20) and cell-free extract of the fungal strain Rhizopus oryzae. The cell-free extract functions as a reducing, shape-directing, as well as stabilizing, agent. Different shapes of gold nanocrystals, for example, triangular, hexagonal, pentagonal, spherical, spheroidal, urchinlike, two-dimensional nanowires, and nanorods, are generated by manipulating key growth parameters, such as gold ion concentration, solution pH, and reaction time. The synthesized nanostructures are characterized by UV/Vis and Fourier-transform infrared spectroscopy, transmission electron microscopy, and energy dispersive X-ray analysis studies. Electron diffraction patterns reveal the crystalline nature of the nanoparticles and a probable mechanism is proposed for the formation of the different structural entities.