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α-galactosidase (EC 3.2.1.22) are glycosidases that catalyze the hydrolysis of α-1,6-linked D-galactosyl residues of different substrates, which has been widely applied in the food industry. Oudemansiella radicata is a kind of precious edible medicinal mushroom, which is a healthy, green, and safe food-derived enzyme source. In this study, a novel acidic α-galactosidase was purified from the dry fruiting bodies of O. radicata by ion-exchange chromatography and gel filtration, and designated as ORG (O. radicata α-galactosidase). ORG was further immobilized to obtain iORG by the sodium alginate–chitosan co-immobilization method. Then, the characterization of free and immobilized enzymes and their potential application in the removal of the RFOs from soymilk were investigated. The results showed that ORG might be a 74 kDa heterodimer, and it exhibited maximum activity at 50 °C and pH 3.0, whereas iORG showed maximum activity at 50 °C and pH 5.5. In addition, iORG exhibited higher thermal stability, pH stability, storage stability, and a better degradation effect on raffinose family oligosaccharides (RFOs) in soymilk than ORG, and iORG completely hydrolyzed RFOs in soymilk at 50 °C within 3 h. Therefore, iORG might be a promising candidate in the food industry due to its excellent stability, high removal efficiency of RFOs from soymilk, and great reusability.

The crude polysaccharides from the fruiting bodies of Clitocybe squamulosa (CSFP) were isolated by hot-water extraction. Two novel polysaccharides, CSFP1-β and CSFP2-α, were further purified by DEAE-52 anion exchange and Sephacryl S-400 gel filtration chromatography, and the purities reached 98.44 and 97.83%, respectively. The structural characteristics and bioactivities of CSFP, CSFP1-β, and CSFP2-α were identified by the combination of chemical and instrumental analysis. Results showed that CSFP was formed by the aggregation of honeycomb spherical materials; CSFP1-β and CSFP2-α were interwoven by reticular and fibrous structures, respectively. Purified components of both CSFP1-β and CSFP2-α showed typical infrared absorption peaks of polysaccharides, and contents of nucleic acid and protein decreased significantly. Simultaneously, CSFP with a molecular weight (Mw) of 1.948 × 104 Da were composed mainly of glucose, mannose, galactose, and rhamnose. CSFP1-β was composed mainly of glucose, galactose, and mannose, while CSFP2-α was composed of glucose, and both their Mw distributions were uneven. Compared with CSFP, the antioxidant activities of CSFP1-β and CSFP2-α were significantly improved (p < 0.05), and they both showed good abilities to bind free cholesterol and bile acid salts in vitro. The binding abilities of the two compounds were found to be 68.62 and 64.43%, and 46.66 and 45.05 mg/g, respectively. CSFP, CSFP1-β, and CSFP2-α had good bacteriostatic effects with a linear increasing relationship to increasing concentration. In addition, CSFP promoted the growth of RAW264.7 cells and has potential immunomodulatory, anti-inflammatory, and anti-tumor activities.

In this study, β-CD@mesoporous SiO2 nanospheres (β-CD@mSi) were prepared by loading β-cyclodextrin (β-CD) onto mesoporous silica nanospheres through an in situ synthesis. This not only solved the defect of β-CD being easily soluble in water, but also changed the physical structure of the mesoporous silica nanospheres. FTIR and XPS results showed that β-CD was successfully loaded onto mesoporous silica nanospheres (mSi), while enhancing the adsorption effect. β-CD@mSi with a monomer diameter of about 150 nm were prepared. At a temperature of 298k, the removal efficiency of a 100 mg/L solution of rifampicin can reach 90% in 4 h and the adsorption capacity was 275.42 mg g−1 at high concentration. Through the calculation and analysis of adsorption kinetics, adsorption isotherms and adsorption thermodynamics based on the experimental data, the reaction is a spontaneous endothermic reaction dominated by chemical adsorption. The electron transfer pathway, structure–activity relationship and energy between β-CD@mSi and rifampicin were investigated by quantum chemical calculations. The accuracy of the characterization test results to judge the adsorption mechanism was verified, to show the process of rifampicin removal by β-CD@mSi more clearly and convincingly. The simulation results show that π–π interaction plays a major interaction in the reaction process, followed by intermolecular hydrogen bonding and electrostatic interactions.

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