Characterisation of polysaccharide from anammox granular sludge and potential application in hydrogel preparation

Microorganisms capable of anaerobic ammonia oxidation (anammox), or the conversion of nitrite and ammonium to dinitrogen, tend to aggregate and form a granular sludge in anammox reactors. This anammox granular sludge is a potential source of polysaccharides due to its richly diverse microbial community and abundant polymers. In this study, anammox polysaccharide (APS) was extracted from anammox granular sludge, and its potential to form hydrogels with alginate was investigated. The yield of APS was 9.91 % ± 0.12 %. The three main monosaccharides in APS were glucose (60.63 % ± 3.45 %), glucuronic acid (13.81 % ± 0.31 %), and rhamnose (18.88 % ± 0.22 %). The antioxidant potential of APS was evaluated through three antioxidant assays, which revealed significant antioxidant benefits at APS concentrations between 100 and 500 mg/L. Furthermore, L929 mouse fibroblasts exhibited high survival rates (>85 %) under different APS concentrations (1-50 μg/mL), indicating the good biological compatibility of APS. A series of hydrogels were prepared by mixing alginate with APS in different ratios (10:0, 9:1, 8:2, 7:3, and 6:4). The swelling ability of the prepared hydrogels in simulated gastric fluid varied between 1.4 and 2.0. In contrast, the swelling ability increased significantly to 10.37 ± 0.01 in simulated intestinal fluid when the ratio of alginate to APS in the hydrogel was 8:2. X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy were also used to analyse the functional groups and specific chemical bonds in the hydrogels. Subsequent loading experiments using bovine serum albumin (BSA) demonstrated that an alginate:APS ratio of 8:2 exhibited the highest loading efficiency for BSA, reaching 80.59 % ± 1.46 %. As the quantity of APS was increased, the release of BSA into simulated gastric fluid was effectively inhibited, with an alginate:APS ratio of 6:4 resulting in the lowest release amount (0.023 % in dry state, 0.11 % in wet state). Overall, this study highlights the derivation of a valuable resource from anammox sludge and offers insights into its potential applications in drug delivery.