Many widely used polymers undergo main chain scission or crosslinking when exposed to radiation. Effects of electron beam irradiation at different doses up to 500 kGy on the biodegradable pullulan polymer films have been investigated by ultraviolet–visible (UV–VIS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and thermogravimetric analysis (TGA) techniques. UV–Visible study shows increase of optical absorbance with increase of doses, attributing to the formation some groups or radicals. FTIR and TGA results reveal the processes of both crosslinking and degradation of polymer taking place depending upon the dose of e-beam radiation. The surface morphology of the film is found to be altered by the e-beam radiation as indicated by SEM micrographs.