Display omitted •Biosynthesis of various PHAs from sludge for different carbon source concentrations.•Correlations among PHAs, EPS and sludge features were evaluated for the first time.•Both EPS and sludge characteristics showed similar trends for each carbon source.•Positive correlations were found between MLSS/polysaccharides and PHAs.•Sludge floc size showed significant negative correlation with PHAs. To understand PHAs accumulation process well in the stand point of the sludge characteristics, polyhydroxyalkanoates (PHAs) accumulation by activated sludge was investigated in correlation with sludge properties, such as particle size, mixed liquor suspended solids (MLSS) and extracellular polymeric substances (EPS) under different carbon sources (e.g., acetate, propionate and mixed carbons) at 2, 4, 6 and 8 g/L. The efficiency of PHAs synthesis from sludge with sodium acetate as carbon source was superior to that with sodium propionate as carbon source and mixed carbon sources in the sequencing biological reactors, while EPS and sludge characteristics showed similar trends for each carbon source. Various carbon sources did not induce significant differences in the correlations. Polyhydroxybutyrate (PHB) showed good correlation with MLSS (R = 0.782) and sludge floc size (R = −0.58). In the reactors with carbon sources of propionate and mixed carbons, polyhydroxyvalerate (PHV) and PHBV correlated with MLSS (R = 0.480 and 0.489, respectively), sludge floc size (R = −0.921 and −0.666, respectively) and concentrations of polysaccharides (R = 0.726, 0.557, respectively). According to the correlations, improving the concentration of sludge, promoting the production of extracellular polymeric substances (EPS) and decreasing the sludge floc size were supposed to be effective approaches for further accumulation of the PHAs. This study is the first to systematically establish the correlations among EPS, characteristics of activated sludge and PHAs accumulation, can provide a theoretical foundation for the engineering design and the operational parameters selection of PHAs production by activated sludge.