The role of hydrazine (N2H4) in anammox metabolism has been widely studied; however, studies on N2H4 biosynthesis by anammox bacteria are limited in the literature. In this context, the current research aims to investigate the enhancement of biological N2H4 production in the anammox process in a long-term manner. The experimental studies started with the optimization of the operating conditions to achieve maximum N2H4 accumulation. Under favorable conditions (pH = 8.97 ± 0.08; T = 35.5 ± 0.5 °C; initial hydroxylamine dose = 1.46 ± 0.01 mM), 17.16 ± 0.64 mg L-1 of N2H4 accumulated in the batch systems. The continuity of N2H4 bioproduction was then evaluated by long-term observations. A continuous flow bioreactor was operated in four consecutive manipulated periods under optimized conditions. In the long-term operated bioreactor, 55.10 ± 0.30 mg L-1 N2H4 was accumulated at optimal conditions, which was 2.5 times higher than reported in the literature. Although manipulation of the bioreactor operating conditions initially resulted in a significant increase in N2H4 bioaccumulation, it subsequently caused a severe deterioration in anammox activity. However, this could be mitigated by increasing the biomass concentration in the anammox systems. In addition, the relative abundance of Candidatus Kuenenia decreased by 1.88% throughout the long-term operation.
Keywords: Anammox metabolism; Bioenergy; Hydrazine; Hydrazine biosynthesis.
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