Study on the mechanism of regulating micromolar Fe utilization and promoting denitrification by guanosine...

A novel multi-signal functional material consisting of Hematin, Fe, and guanosine monophosphate (GMP) was successfully constructed (Hematin@Fe/GMP) to enhance denitrification efficiency based on the signal network regulation of electron transfer, micromolar Fe utilization, and microbial community. Hematin@Fe/GMP enhanced nitrate reduction rate by 2.33-fold with a 9.9 mg L^-1 h^-1 reduction rate. The mechanisms of accelerated denitrification were elaborated deeply from the electrochemical experiments, microbial metabolism activity, key enzyme activity, gene expression, and microbial community. Specifically, electrochemical experiments and X-ray photoelectron spectroscopy demonstrated that the released redox signal (Fe²⁺/Fe³⁺) promoted the increased redox substances (extracellular polymeric substances, cytochrome c, and riboflavin) to accelerate electron transfer efficiency. Metagenomic analysis suggested the released Fe utilization signal modulated siderophores genes (fhuB, fhuC, and fhuD) to promote the uptake and utilization of micromolar Fe, which was more conducive to synthesizing cytochrome c. Moreover, extracellular polymeric substances (EPS) stripping experiments demonstrated that the membrane-anchored cyt-c could shuttle in EPS and bind with Hematin@Fe/GMP to form an electrical conduit for accelerating denitrification efficiency. In inhibition experiments, Hematin@Fe/GMP could break down electron transfer barriers and restore/compensate for the electron transfer chain. Meanwhile, Hematin@Fe/GMP could restore the electrical signal disruption and synergize with the enriched signaling-capable microorganisms (Stutzerimonas and Thauera) to regulate quorum sensing. This research introduced multi-signal modulation of Hematin@Fe/GMP on denitrification and provided strategies for accelerating the biological transformation process and effectively utilizing micromolar Fe in practical applications.