In this research, two sequencing batch reactors (R1 and R2) were operated with different feeding strategies to investigate the effects of elongated anaerobic feeding mode on the granulation process. For this purpose, R1 was operated in a short-feed strategy (5 min) as a reference, whereas an extended anaerobic feeding of 85 min was applied in R2. Results showed that aerobic granules formed in R1 were denser and more uniform with lower sludge volume index than those formed in R2. Investigation of tightly bound extracellular polymeric substances (TB-EPS) showed that aerobic granules in R1 produced lower amounts of TB-EPS than those in R2. This was due to the bigger and more compact granules cultivated in R1 in comparison with looser structure granules in R2 with higher ﬂocculent sludge percentage. The relative hydrophobicity of granules in both reactors was increased with culture time and reached about 63 and 65% at day 70 for R1 and R2, respectively. Moreover, no significant correlation between protein/polysaccharide ratio and hydrophobicity was found, which showed that protein in loosely bound extracellular polymer substances was not the predominant hydrophobic component. Considering the outcomes of this study, it can be concluded that the elongated anaerobic feeding strategy was unfavourable for cultivating aerobic granules.