Magnetite can be considered as an iron-rich carrier particles that can be ionized into Fe2+ and Fe3+ which improves the activity and aggregation of anammox bacteria. Three samples from this carrier assisted granulation reactor with size groups including Flocs, FL (0-300 µm), Small Granules, SG (300-500 µm) and Large Granules, LG (500-1000 µm) were used in this study. It was observed that as the granule size increased, the iron-rich carrier content increased, and their active crystals improved the microbial cell density. Specific anammox activity (SAA) was 34.63 ± 5.02, 55.29 ± 5.14, and 63.81 ± 7.50 mg-N/g-VSS/d for FL, SG and LG, respectively. In addition, in heme c content of LG was 31.5 % higher than SG and 62.9 % higher than FL. An in-depth study into the extracellular polymeric substances (EPS) showed that the secretion intensity of essential proteins followed the order of FL < SG < LG in loosely bound EPS and FL > SG > LG in tightly bound EPS. Functional group analysis confirmed that the hydrophobic CN and NH stretching vibration band had almost 3.5 times higher transmittance intensity in LG than the other sizes and the corresponding ratio of α-helix/(β sheet + random coil) in secondary derivative proteins analysis showed tightness in the protein structures of FL. The relative abundance of Brocadia Sinica increased from 0 % in FL to a high of 20.46 % in LG. This study aims to communicate the essence of in-depth EPS analysis beyond the usual EPS yield and major contents of proteins (PN) and polysaccharides (PS) analysis.
Keywords: Agglomeration; Anammox; EPS; Granule size; Hydrophilicity; Hydrophobicity.
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