Functional microorganisms and enzymes related nitrogen cycle in the biofilm performing simultaneous nitrification and denitrification

Simultaneous nitrification and denitrification (SND) is a potential energy-saving process in wastewater treatment while the nitrogen removal mechanism is still unclear due to the lack of information about the functional microbes and enzymes. Sequencing batch biofilm reactors were implemented to achieve efficient SND. Eight nitrogen removal related microorganisms out of the top abundant 20 microbial community and reference species were used to construct a phylogenetic tree. Functional enzymes and modules analysis were investigated to reveal the SND pathway: in the aerobic part of the biofilm, ammonia oxidation was catalyzed by complete ammonia oxidizers while in the inner anoxic part, denitrification, dissimilatory nitrate reduction (DNRA) and nitrogen fixation (NF) cooperated to stimulate nitrate removal. These results provide a practical aeration control strategy to achieve SND and indicate that DNRA and NF are important nitrogen removal pathways that should not be ignored in the SND mechanism.