Biogranulation is an effective biological technology suitable for the treatment of various wastewaters. However, the major drawback of this technique is the long start-up period for biogranule development. Hence, the primary focus of this study was on cell surface hydrophobicity which is the main parameter that indicates cell agglomeration during the initial self-immobilization process of aerobic granulation. The effects of sludge concentration and magnetic activated carbon on cell surface hydrophobicity were investigated in this study. Response surface methodology (RSM) was applied to design, analyze, and optimize the outcome of the study. Experiments were performed at sludge concentration of 1,000-3,000mg/L and magnetic activated carbon mass of 1-5g/L with 24hr of aeration time. The results show that both variables yielded a positive significant effect on the initial development of aerobic granulation with 56% surface hydrophobicity. Interaction effects between variables on the responses were significant with positive estimated interaction effect at all different measured aeration time. The magnetic activated carbon acted as nuclei to induce bacterial attachment and further enhanced the initial process of biogranule development under optimal condition of 1:1.1 (sludge concentration: magnetic activated carbon). PRACTITIONER POINTS: Cell surface hydrophobicity was evaluated Magnetic activated carbon enhanced cell surface hydrophobicity Response surface methodology was employed for analyses Magnetic activate carbon mass and biomass concentration was significant Magnetic activated carbon acted as nuclei to improve biogranulation.