Promotion of anammox process by different graphene-based materials: Roles of particle size and oxidation degree

Graphene oxide (GO) and reduced graphene oxide (RGO) have been applied in the anaerobic ammonium oxidation (anammox) process for nitrogen removal as electron shuttles. However, there is still controversy about their efficacy. In this study, nine graphene-based materials with a gradient of three particle sizes (large (l), medium (m) and small (s) sizes) and oxidation degrees, were used to compare their effects on the anammox process efficiency. The graphene-based materials include GO and its reduced products (RGO250 and RGO800) obtained at temperatures of 250 °C and 800 °C respectively. It was observed that their enhancements on the anammox process were in the order of GO > RGO800 > RGO250. In detail, at the dose of 100 mg/L, specific anammox activities (SAA) were promoted by 6.7% (l-GO), 4.9% (l-RGO800), 11.5% (m-GO), 7.3% (m-RGO800), 13.2% (s-GO) and 8.3% (s-RGO800) compared to the control respectively; while RGO250 with the same dose inhibited the process. In addition, the enhancement of the anammox process was increasing with the decreasing size of GO and RGO800. The nitrite reductase (NIR) activity was greatly increased by up to 24.9% with the presence of GO, which might be attributed to organized and specific electron transport with oxygen functional groups. The finding of hydroxyl on RGO and increasing content of oxygen determined after reaction detected by Fourier transform infrared spectroscopy and energy dispersive spectrometer respectively, indicated the essential condition for RGO's function on transferring electrons for key enzymes in annamox bacteria. Most importantly, O/C (Oxygen/Carbon) ratios of graphene-based materials had greater effects on the promotion of the anammox process than the particle size and electrical conductivity.