Interspecies cooperation-driven photogenerated electron transfer processes and efficient multi-pathway nitrogen removal in the g-C 3 N 4-anammox consortia biohybrid system

Photocatalytic materials-microbial biohybrid systems pave the way for solar-driven wastewater nitrogen removal. In this study, interspecies cooperation in photogenerated electron transfer and efficient nitrogen removal mechanism in the g-C 3 N 4 -anammox consortia biohybrid system were first deciphered. The results indicated that the essential extracellular electron carriers (cytochrome c and flavin) for anammox genomes were provided by associated bacteria (BACT3 and CHLO2). This cooperation, regulated by the ArcAB system and electron transfer flavoprotein, made anammox bacteria the primary photogenerated electron sink. Furthermore, an efficient photogenerated electron harness was used to construct a reductive glycine pathway (rGlyP) in anammox bacteria inventively, which coexisted with the Wood - Ljungdahl pathway (WLP), constituting a dualpathway carbon fixation model, rGlyP-WLP. Carbon fixation products efficiently contributed to the tricarboxylic acid cycle, while inhibiting electron diversion in anabolism. Photogenerated electrons were targeted channeled into nitrogen metabolism-available electron carriers, enhancing anammox and dissimilatory nitrate reduction to ammonium (DNRA) processes. Moreover, ammonia assimilation by the glycine cleavage system in rGlyP established an alternative ammonia removal route. Ultimately, multi-pathway nitrogen removal involving anammox, DNRA, and rGlyP achieved 100 % ammonia removal and 94.25 % total nitrogen removal efficiency. This study has expanded understanding of anammox metabolic diversity, enhancing its potential application in carbon-neutral wastewater treatment.