Currently, there is little comparative data on the colloidal stability and the toxicity of ultraviolet (UV)- and visible-light (VL)-transformed graphene oxide (GO). In order to identify this knowledge gap, the physicochemical properties of UV/VL-transformed GO are investigated in detail. Attempts are made to correlate the physicochemical alterations of UV/VL-transformed GO to the observed changes in its colloidal properties and toxicity. The results show that both UV and VL irradiations induce the significant change in the color, UV-vis absorbance, morphology, surface charge, size, oxygen containing functional groups, total of carbon, and photoluminescence properties of GO. The photo-reaction behavior of GO under UV exposure is different from that under VL irradiation in terms of reaction rate, order, and extent. Finally, the UV and VL irradiations show different effects not only on the colloidal stability of GO in the City water and Dongpu Lake water, but also on the toxicity of GO to Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria. This study clearly shows how the environmental fate and risk of GO are modified by UV and VL irradiations.