The effect of distribution pattern of polysaccharides (PS) and proteins (PN) in activated sludge (AS) stratification with different aeration rates on membrane fouling and rejection efficiency were investigated. During high aeration, PN and PS concentrations increased in supernatant, the dominant fraction (84% of PN and 73% of PS) was small molecules (<1 kDa). Less slime and loose bound extracellular polymeric substances (LB-EPS), more tight bound EPS (TB-EPS) were observed compared with low aeration. The decrease in PN/PS ratio and Ca2+ concentration within EPS deteriorated AS flocculation ability. At slow trans-membrane pressure (TMP) rise stage, fouling rate under high aeration was 41% lower than low aeration due to lower PN within EPS outer. Low PS rejection rate (about 23%) leaded to higher PS in effluent at this stage. High PS rejection rate (about 94%) at rapid TMP rise stage resulted in about 2.2-time higher fouling rate than that low aeration. (C) 2015 Elsevier Ltd. All rights reserved.