Alcaligenes faecalis C16 was found to have the ability to heterotrophically nitrify and aerobically denitrify. In order to further understand its nitrogen removal ability and mechanism, the growth and ammonium removal response were investigated at different C/N ratios and ammonium concentrations in the medium with citrate and acetate as carbon source separately. Furthermore, experiments of nitrogen sources, production of nitrogen gas and enzyme assay were conducted. Results show that the bacterium converts NH4+-N and produces NH2OH during the growing phase and nitrite accumulation is its distinct metabolic feature. A. faecalis C16 is able to tolerate not only high ammonium concentration but also high C/N ratio, and the ammonium tolerance is associated with carbon source and C/N ratio. The nitrogen balance under different conditions shows that approximately 28%-45% of the initial ammonium is assimilated into the cells, 44%-60% is denitrified and several percent is converted to nitrification products. A. faecalis C16 cannot utilize hydroxylamine, nitrite or nitrate as the sole nitrogen source for growth. However, nitrate can be used when ammonium is simultaneously present in the medium. A possible pathway for nitrogen removal by C16 is suggested. The preliminary enzyme assay provides more evidence for this nitrogen removal pathway. (C) 2015 The Chemical Industry and Engineering Society of China, and Chemical Industry Press. All rights reserved.