Periphytic biofilms are microbial aggregates commonly present in submerged aquatic environments and play a significant role in nutrient cycling. In recent years, utilization of natural periphytic biofilms in wastewater treatment and water restoration attracts growing research interests. Light and temperature are two important environmental factors known to affect the development of periphytic biofilms and can be manipulated for the regulation of the biofilm properties. In this work, effects of light and temperature on the development and function (denitrification potential) of periphytic biofilms were investigated using a microcosm experiment. Results showed that thicker periphytic biofilms with higher Chlorophyll a, extracellular polymeric substances (EPS), and total phosphorus contents were developed under higher temperature. Whereas, biomass accumulation was more rapid for periphytic biofilms under higher irradiance. The denitrification potential rate was negatively associated with irradiance, which can be linked to the influence of irradiance on biofilm structure and microbial composition. A relatively lower irradiance is recommended when using periphytic biofilms in nitrogen removal from wastewater. (C) 2017 Elsevier B.V. All rights reserved.