An anaerobic sequencing batch reactor(ASBR) inoculated with activated sludge was employed to investigate the start-up of anaerobic ammonium oxidation(ANAMMOX) process and the spatial distribution of extracellular polymeric substances(EPS) in ANAMMOX granules. The results showed that the removal rates of NH4+-N and NO2--N reached more than 99%. The removal rate of total nitrogen(TN) was 89.87%±0.43% and the TN removal load in the ASBR was 1.7 kg·(m3·d)-1. The ratios of nitrite consumption and nitrate production to ammonium consumption were 1.32±0.08 and 0.24±0.03, respectively. Besides, the pH and the effluent nitrate concentration in bulk liquid were considered as two simple indicators for rapid diagnosis of ANAMMOX performance. The proteins(PN) was the main component of EPS in ANAMMOX granular sludge. The PN and polysaccharides(PS) in ANAMMOX granules were (59.61±5.64) mg·g-1 and (12.21±2.04) mg·g-1, respectively, with the ratio of PN/PS reaching approximately 4.88±1.39. β-D-glucofuranose and dead cells were distributed in the outermost layer of granules, while living cells, PN, lipids, α-glucosamine and α-mannose were distributed throughout the granules and mainly concentrated on the outside of granules. PN and lipids constituted the framework of ANAMMOX granules and ANAMMOX bacteria were embedded among the PN and lipids matrix.