•Higher NH4+-N load led to higher oxygen transfer rate and lower NOB activity.•NOB was inhibited by low DO concentration in the limited space of aerobic biofilm.•Closed-end aeration mode favored Nitrosomonas and inhibited Nitrospira.•A strategy was proposed for quick start-up and stable mainstream PN in MABR. The proliferation of nitrite oxidizing bacteria (NOB) still remains as a major challenge for nitrogen removal in mainstream wastewater treatment process based on partial nitrification (PN). This study investigated different operational conditions to establish mainstream PN for the fast start-up of membrane aerated biofilm reactor (MABR) systems. Different oxygen controlling strategies were adopted by employing different influent NH4+-N loads and oxygen supply strategies to inhibit NOB. We indicated the essential for NOB suppression was to reduce the oxygen concentration of the inner biofilm and the thickness of aerobic biofilm. A higher NH4+-N load (7.4 g-N/(m2·d)) induced higher oxygen utilization rate (14.4 g-O2/(m2·d)) and steeper gradient of oxygen concentration, which reduced the thickness of aerobic biofilm. Employing closed-end oxygen supply mode exhibited the minimum concentration of oxygen to realize PN, which was over 46% reduction of the normal open-end oxygen mode. Under the conditions of high NH4+-N load and closed-end oxygen supply mode, the microbial community exhibited a comparative advantage of ammonium oxidizing bacteria over NOB in the aerobic biofilm, with a relative abundance of Nitrosomonas of 30–40% and no detection of Nitrospira. The optimal fast start-up strategy was proposed with open-end aeration mode in the first 10 days and closed-end mode subsequently under high NH4+-N load. The results revealed the mechanism of NOB inhibition on the biofilm and provided strategies for a quick start-up and stable mainstream PN simultaneously, which poses great significance for the future application of MABR. Display omitted