The study investigated the effect of acetate/starch mixture on the formation of storage biopolymers as compared with the storage mechanism in systems fed with these compounds as single substrates. Experiments involved two sequencing batch reactor sets operated at steady state, at sludge ages of 8 and 2 days, respectively. Each set included three different runs, one fed with acetate, the other with starch and the last one with the acetate/starch mixture. In single substrate systems with pulse feeding, starch was fully converted to glycogen, whereas 25 % of acetate was utilized for direct microbial growth at sludge age of 8 days, together with polyhydroxybutyric acid (PHB) storage. The lower sludge age slightly increased this fraction to 30 %. Feeding of acetate/starch mixture induced a significant increase in acetate utilization for direct microbial growth; acetate fraction converted to PHB dropped down to 58 and 50 % at sludge ages of 8 and 2 days respectively, while starch remained fully converted to glycogen for both operating conditions. Parallel microbiological analyses based on FISH methodology confirmed that the biomass acclimated to the substrate mixture sustained microbial fractions capable of performing both glycogen and PHB storage.