Phototrophic biofilms are exposed to multiple stressors that can affect them both directly and indirectly. By modifying either the composition of the community or the physiology of the microorganisms, press stressors may indirectly impact the ability of the biofilms to cope with disturbances. Extracellular polymeric substances (EPS) produced by the biofilm are known to play an important role in its resilience to various stresses. The aim of this study was to decipher to what extent slight modifications of environmental conditions could alter the resilience of phototrophic biofilm EPS to a realistic sequential disturbance (4-day copper exposure followed by a 14-day dry period). By using very simplified biofilms with a single algal strain, we focused solely on physiological effects. The biofilms, composed by the non-axenic strains of a green alga ( Uronema confervicolum ) or a diatom ( Nitzschia palea ) were grown in artificial channels in six different conditions of light intensity, temperature and phosphorous concentration. EPS quantity (total organic carbon) and quality (ratio protein/polysaccharide, PN/PS) were measured before and at the end of the disturbance, and after a 14-day rewetting period. The diatom biofilm accumulated more biomass at the highest temperature, with lower EPS content and lower PN/PS ratio while green alga biofilm accumulated more biomass at the highest light condition with lower EPS content and lower PN/PS ratio. Temperature, light intensity, and P concentration significantly modified the resistance and/or recovery of EPS quality and quantity, differently for the two biofilms. An increase in light intensity, which had effect neither on the diatom biofilm growth nor on EPS production before disturbance, increased the resistance of EPS quantity and the resilience of EPS quality. These results emphasize the importance of considering the modulation of community resilience ability by environmental conditions, which remains scarce in the literature.