Membrane fouling precludes the widespread application of membrane filtration system from treating wastewater and drinking water, and occurs even under sub-critical flux operations. Hence the characteristics and behavior of membrane foulants should be thoroughly investigated, so as to find ways to reduce membrane fouling in membrane bioreactors. The purpose of this study is to compare the membrane fouling potential at different sub-critical flux operations and for different hydrophobic/hydrophilic membranes, and to investigate the vertical distribution of membrane foulants in a cake layer. Results showed that higher fouling propensity which occurred under 80% of critical flux of hydrophilic membrane was associated with the soluble fraction of proteins and polysaccharides, compared with 60% of critical flux. The cell-bound components were dominant under hydrophobic membrane operation. The highest concentration of proteins and polysaccharides was found between 40% and 80% of the depth of the cake layer. (C) 2014 Elsevier Ltd. All rights reserved.