A laboratory-scale submerged membrane bioreactor (SMBR) with a hollow-fibre membrane module was used to investigate membrane fouling under various operational conditions. The results showed that the sludge supernatant inside the SMBR had a consistently higher organic content than the MBR effluent. Detailed analysis revealed a pool of organic substances, classified as biopolymer clusters (BPC), in the SMBR system that were larger in size than the soluble microbial products (SMP). The BPC content in the MBR sludge mixture ranged from 0.7 to 18.8 mg/L in terms of the total organic carbon (TOC), with an average of 5.6+/-3.5mg/L, which was about twice the SMP concentration in the suspension. Under a fluorescent microscope and using DAPI staining, the BPC in the sludge supernatant after centrifugation were found to be particulate organic substances that were independent of the sludge flocs and had a size distribution up to 50 microm. The findings of the experiment suggest that BPC are an important foulant and have a profound effect on membrane fouling. The fouling rate in the reactor, as indicated by the increase in trans-membrane pressure (TMP), correlates to a certain extent with the BPC concentration in the sludge suspension under various conditions. It is argued that BPC are a special form of organic substances that are formed by the adsorption and affinity clustering of mainly SMP within the sludge layer deposited on the membrane surface. The BPC can be detached with the sludge from the membrane and returned to the suspension. The accumulation of BPC in the SMBR sludge mixture would facilitate the formation of a sludge fouling layer on the membrane surface, thus causing a serious fouling problem.