Anammox Coupled with Photocatalyst for Enhanced Nitrogen Removal and the Activated Aerobic Respiration of Anammox Bacteria Based on cbb3-Type Cytochrome c Oxidase

Abstract

This study introduced photogenerated electrons into the anammox system by coupling them to a g-C3N4 nanoparticle photocatalyst. A high nitrogen removal efficiency (94.25%) was achieved, exceeding the biochemical limit of 89% imposed by anammox stoichiometry. Photogenerated electrons boosted anammox metabolic activity by empowering key enzymes (NIR, HZS, and WLP-related proteins) and triggered rapid algal enrichment by enhancing the algal Calvin cycle, thus developing multiple anammox–algae synergistic nitrogen removal processes. Remarkably, the homologous expression of cbb3-type cytochrome c oxidase (CcO) in anammox bacteria was discovered and reported in this study for the first time. This conferred aerobic respiration capability to anammox bacteria and rendered them the principal oxygen consumer under 7.9–19.8 mg/L dissolved oxygen, originating from algal photosynthesis. Additionally, photogenerated electrons selectively targeted the cb1 complex and cbb3-type CcO as activation sites while mobilizing the RegA/B regulatory system to activate the expression of cbb3-type CcO. Furthermore, cbb3-type CcO blocked oxidative stress in anammox by depleting intracellular oxygen, a substrate for reactive oxygen species synthesis. This optimized the environmental sensitivity of anammox bacteria and maintained their high metabolic activity. This study expands our understanding of the physiological aptitudes of anammox bacteria and provides valuable insights into applying solar energy for enhanced wastewater treatment.