Purification and structural characterization of a neutral polysaccharide from Boletus auripes using self-made quaternary chitosan cryogel.

The purification of polysaccharides is an essential preliminary step in determining their chemical structure, although it presents significant challenges. In this research, a macro-porous monolith of quaternary chitosan cryogel was synthesized for the purification of a neutral polysaccharide from Boletus auripes. A homogeneous neutral polysaccharide (BAP-1a1) with a weight-average molecular weight of 4.13 × 10 Da and a polydispersity index of 1.28 was successfully isolated. The structure of BAP-1a1 was elucidated through a comprehensive characterization utilizing size exclusion chromatography (SEC) combined with laser light scattering (LLS), infrared spectroscopy, monosaccharide composition analysis, methylation analysis, and nuclear magnetic resonance (NMR) spectroscopy. The results revealed that the BAP-1a1 was characterized as a glucan with a backbone structure consisting of 1,4-α-D-Glcp and 1,3-β-D-Glcp glycosidic linkages in a molar ratio of 2:1. Additionally, a minority of branched chains of 1-α-D-Glcp are attached to 1,3-β-D-Glcp residues at the C6 position. In vitro antioxidant activity assays demonstrated that BAP-1a1 exhibits a dose-dependent scavenging effect on ABTS and DPPH radicals with EC values of 0.58 and 1.04 mg/mL, respectively. These findings indicated that Boletus auripes possesses the potential to be utilized as a natural agent in antioxidant functional foods.