Structure-activity relationship of low molecular weight Astragalus membranaceus polysaccharides produced by Bacteroides.

Astragalus membranaceus polysaccharides (APS) possess significant biological activities, such as anti-tumor, antiviral, and immunomodulatory activities. However, there is still a lack of research on the structure-activity relationship of APS. In this paper, two carbohydrate-active enzymes from Bacteroides in living organisms were used to prepare degradation products. The degradation products were divided into APS-A1, APS-G1, APS-G2, and APS-G3 according to molecular weight. Structural analysis showed that all degradation products had an α-1,4-linked glucose backbone, but APS-A1 and APS-G3 also had branched chains of α-1,6-linked galactose or arabinogalacto-oligosaccharide. In vitro, immunomodulatory activity evaluation results indicated that APS-A1 and APS-G3 had better immunomodulatory activity, while the immunomodulatory activities of APS-G1 and APS-G2 were comparatively weaker. Molecular interaction detection showed that APS-A1 and APS-G3 could bind to toll-like receptors-4 (TLR-4) with a binding constant of 4.6 × 10-5 and 9.4 × 10-6, respectively, while APS-G1 and APS-G2 failed to bind to TLR-4. Therefore, the branched chains of galactose or arabinogalacto-oligosaccharide played a crucial role in the immunomodulatory activity of APS.

Structure and function characterization of the α-L-arabinofuranosidase from the white-rot fungus Trametes hirsuta.

α-L-Arabinofuranosidases (Abfs) play a crucial role in the degradation of hemicelluloses, especially arabinoxylans (AX). Most of the available characterized Abfs are from bacteria, while fungi, as natural decomposers, contain Abfs with little attention given. An arabinofuranosidase (ThAbf1), belonging to the glycoside hydrolase 51 (GH51) family, from the genome of the white-rot fungus Trametes hirsuta, was recombinantly expressed, characterized, and functionally determined. The general biochemical properties showed that the optimal conditions for ThAbf1 were pH 6.0 and 50°C. In substrate kinetics assays, ThAbf1 preferred small fragment arabinoxylo-oligosaccharides (AXOS) and could surprisingly hydrolyze di-substituted 23,33-di-L-arabinofuranosyl-xylotriose (A2,3XX). It also synergized with commercial xylanase (XYL) and increased the saccharification efficiency of arabinoxylan. The crystal structure of ThAbf1 indicated the presence of an adjacent cavity next to the catalytic pocket which led to the ability of ThAbf1 to degrade di-substituted AXOS. The narrow binding pocket prevents ThAbf1 from binding larger substrates. These findings have strengthened our understanding of the catalytic mechanism of GH51 family Abfs and provided a theoretical foundation for the development of more efficient and versatile Abfs to accelerate the degradation and biotransformation of hemicellulose in biomass. KEY POINTS: • ThAbf1 from Trametes hirsuta degraded di-substituted arabinoxylo-oligosaccharide. • ThAbf1 performed detailed biochemical characterization and kinetics. • ThAbf1 structure has been obtained to illustrate the substrate specificity.