Comparison of physicochemical and biochemical properties of natural and arginine-modified melanin from medicinal mushroom Ganoderma lucidum.

Melanin is a hydrophobic biomolecule produced widely in fungi. Compared with other fungi, health benefits have been associated with medicinal mushrooms, which may provide an excellent source of natural melanin. Nevertheless, the hydrophobicity of melanin may limit its applications. Consequently, the present study was carried out on isolation of melanin from the medicinal mushroom Ganoderma lucidum (GLM) and modification with arginine to improve its solubility. The physicochemical and biochemical properties of melanin were evaluated including structural characterization, solubility, stability, antioxidant activities, and inhibitory effect on pancreatic lipase activity. Arginine-modified melanin showed better solubility, higher color value, stronger antioxidant activity, and stronger inhibitory effect on pancreatic lipase activity in vitro than GLM. In addition, both have good stability in the dark and natural light. These results opened possibilities for providing an excellent source of natural melanin in health food or food additives fields.

Hydrolysis of oligosaccharides by a fungal α-galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor.

A novel acidic α-galactosidase (EC 3.2.1.22) designated as Leucopaxillus tricolor α-galactosidase (LTG) has been purified to homogeneity from the fruiting bodies of L. tricolor to 855-fold with a specific activity of 956 U mg-1 by the application of chromatography and gel filtration. The molecular mass of LTG was estimated to be 60 kDa as determined by both SDS-PAGE and by gel filtration. The purified enzyme was identified by LC-MS/MS and four inner amino acid sequences were obtained. When 4-nitrophenyl α-D-glucopyranoside (pNPGal) was used as substrate, the optimal pH and optimal temperature of LTG were pH 5.0 and 50 °C, respectively. The enzyme activity was strongly inhibited by Hg2+ , Fe3 , Cu2+ , Cd2+ , and Mn2+ ions. The chemical modification agent N-bromosuccinimide (NBS) completely inhibited the enzyme activity of LTG, indicating the paramount importance of tryptophan residue(s) to its enzymatic activity. Besides, LTG displayed wide substrate diversity with activity toward a variety of substrates such as stachyose, raffinose, melibiose, locust bean gum, and guar gum. Given the good ability of degrading the non-digestible and flatulence-causing oligosaccharides, this fungus may become a useful source of α-galactosidase for multiple applications.

Characterization of Polysaccharides with Antioxidant and Hepatoprotective Activities from the Edible Mushroom Oudemansiella radicata.

The preliminary structure, in vitro antioxidant and in vivo hepatoprotective activities of water-soluble polysaccharides (ORWP) and alkali-soluble polysaccharides (ORAP), prepared from the mushroom Oudemansiella radicata, were investigated. Both ORWP and ORAP were heteropolysaccharides with mannose, glucose and galactose being the main monosaccharide components. Regarding the antioxidant activities, ORWP and ORAP showed effective 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, hydrogen peroxide scavenging activity and lipid peroxidation inhibitory effects, as well as moderate reducing power and Fe2+ chelating activity. For the hepatoprotective activity, administration of ORWP and ORAP prevented the increase in serum alanine aminotransferase and aspartate aminotransferase activities in a carbon tetrachloride-induced acute liver damage model, suppressed hepatic malondialdehyde formation and stimulated the activities of hepatic superoxide dismutase and glutathione peroxidase. Thus, we speculate that ORWP and ORAP may protect the liver from CCl4-induced hepatic damage via antioxidant mechanisms.

A Fungal α-Galactosidase from Tricholoma matsutake with Broad Substrate Specificity and Good Hydrolytic Activity on Raffinose Family Oligosaccharides.

An acidic α-galactosidase designated as TMG was purified from the fruiting bodies The purification protocol entailed ion exchange chromatography on Q-Sepharose and of Tricholoma matsutake with 136-fold purification and a specific activity of 909 units/mg. Mono-Q and fast protein liquid chromatography on Superdex 75. TMG is a monomeric protein exhibiting a molecular mass of 47 kDa in SDS-PAGE and gel filtration. The purified enzyme was identified by LC-MS/MS and three inner amino acid sequences were obtained. The optimum pH and temperature for TMG with pNPGal as substrate were pH 4.5 and 55 °C, respectively. The α-galactosidase activity was strongly inhibited by K+, Ca2+, Cd2+, Hg2+, Ag+ and Zn2+ ions. The enzyme activity was inhibited by the chemical modification agent N-bromosuccinimide (NBS), indicating the importance of tryptophan residue(s) at or near the active site. Besides hydrolyzing pNPGal, TMG also efficaciously catalyzed the degradation of natural substrates such as stachyose, raffinose, and melibiose. Thus TMG can be exploited commercially for improving the nutritional value of soy milk by degradation of indigestible oligosaccharides.

Isolation of an Angiotensin I-Converting Enzyme Inhibitory Protein with Antihypertensive Effect in Spontaneously Hypertensive Rats from the Edible Wild Mushroom Leucopaxillus tricolor.

An 86-kDa homodimeric angiotensin I-converting enzyme (ACE) inhibitory protein designated as LTP was isolated from fruit bodies of the mushroom Leucopaxillus tricolor. The isolation procedure involved ultrafiltration through a membrane with a molecular weight cutoff of 10-kDa, ion exchange chromatography on Q-Sepharose, and finally fast protein liquid chromatography-gel filtration on Superdex 75. LTP exhibited an IC50 value of 1.64 mg∙mL-1 for its ACE inhibitory activity. The unique N-terminal amino acid sequence of LTP was disclosed by Edman degradation to be DGPTMHRQAVADFKQ. In addition, seven internal sequences of LTP were elucidated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results of the Lineweaver-Burk plot suggested that LTP competitively inhibited ACE. Both LTP and the water extract of L. tricolor exhibited a clear antihypertensive effect on spontaneously hypertensive rats.

Isolation and characterization of a novel lectin from the edible mushroom Stropharia rugosoannulata.

To date, only a few steroids have been isolated from the mushroom Stropharia rugosoannulata which can be cultivated. In this paper, a novel lectin (SRL) with a molecular weight of 38 kDa, and a unique IKSGVYRIVSWQGALGPEAR N-terminal sequence was isolated from S. rugosoannulata, which represents the first protein isolated from the mushroom. The purification methods included (NH4)2SO4 precipitation, ion exchange chromatography on CM-cellulose, Q-Sepharose, and SP-Sepharose, and gel- filtration on Superdex-75. The lectin was adsorbed on all three types of ion exchangers and was purified more than 450-fold. The lectin was stable below 70 °C (with half of the activity preserved at 80 °C), and in the presence of NaOH and HCl solutions up to a concentration of 12.5 mM and 25 mM, respectively. The hemagglutinating activity of SRL was inhibited by inulin. Cd2+ and Hg2+ ions strongly reduced the hemagglutinating activity at concentrations from 1.25 mM to 10 mM. SRL exhibited anti-proliferative activity toward both hepatoma Hep G2 cells and leukemia L1210 cells, with an IC50 of 7 µM and 19 µM, respectively. The activity of HIV-1 reverse transcriptase could also be inhibited by SRL, with an IC50 of 10 µM.

Functional Properties, Rheological Characteristics, Simulated Digestion, and Fermentation by Human Fecal Microbiota of Polysaccharide from Morchella importuna.

Morchella importuna polysaccharide (MIP) has been proven to have obvious hypoglycemic effects on mice with type 2 diabetes (T2DM). This study looked at the functional and rheological characteristics of MIP, and investigated the effects of MIP on the human fecal microbiota through in vitro fermentation experiments. The outcomes demonstrate the excellent oil-holding capacity, emulsifying, foaming, and rheological characteristics of MIP. After salivary gastrointestinal digestion, the Mw of MIP decreased from 398.2 kDa and 21.5 kDa to 21.9 kDa and 11.7 kDa. By 16S rRNA sequencing of bacteria fermented in vitro, it was found that MIP did not improve the richness and diversity of intestinal microorganisms, but it may exert an anti-T2DM function by significantly increasing the relative abundance of Firmicutes and promoting Ruminococcaceae_UCG_014, Bacteroides, and Blautia proliferation. Escherichia-Shigella could also be inhibited to improve the intestinal microenvironment. In addition, the fermentation of MIP increased the total short-chain fatty acid (SCFA) concentration from 3.23 mmol/L to 39.12 mmol/L, and the propionic acid content increased significantly. In summary, MIP has excellent processing performance and is expected to exert potential anti-T2DM activity through the human intestinal microbiota, which has broad market prospects.

Polysaccharide from Clitocybe squamulosa: Gel-forming properties and its compound effect with food thickener.

To study the gel-forming properties of polysaccharide from the fruiting body of Clitocybe squamulosa (CSFP) and its degradation product (UH-CSFP), the changes in steady-state and dynamic rheological properties of CSFP and UH-CSFP under different conditions (polysaccharide mass fraction, temperature, pH, and salt ion concentration) were studied. Polysaccharides with good gel-forming properties were selected and mixed with common edible thickeners (gelatin, guar gum, and locust bean gum), after which the properties of the composite gel were assessed. The steady-state rheological results showed that CSFP and UH-CSFP were pseudoplastic fluids, their apparent viscosity decreased with increasing temperature, the viscosity was greatest when the pH was 7. The addition of Na+ and Ca2+ could increase the viscosity, and the viscosity of UH-CSFP was lower than that of CSFP at the same mass fraction. The results of dynamic rheology indicated that G´ and G´´ of CSFP and UH-CSFP increased with increasing mass fraction, pH, and ion concentration (0.01 M to 1 M), and G´´ was always smaller than G´ indicating weak gel behavior. The thixotropy-related experimental results showed that the thixotropy ring area of CSFP and UH-CSFP increased with increasing mass fraction, the ring area of CSFP was larger than that of UH-CSFP, and the gel strength of CSFP was greater than that of UH-CSFP. The results of CSFP and three types of edible gels showed that the composite gels were pseudoplastic fluids, and their apparent viscosity was ranked (in descending order) as follows: guar bean gum, locust bean gum, and gelatin. The addition of CSFP improved the gel-forming properties of guar gum but did not significantly improve the gel properties of locust bean gum and gelatin. This study provides a theoretical basis for the selection of processing methods and the application of polysaccharides.

The effect of sequential extraction on the physicochemical and rheological properties of Naematelia aurantialba polysaccharides.

To overcome the difficulty of separation and low rate of extraction caused by highly viscous polysaccharides from Naematelia aurantialba (NA), four N. aurantialba polysaccharides (NAPs) were sequentially extracted using water (enzyme-/ultrasound-assisted extraction), alkali (0.1 mol/L NaOH), and acid (0.1 mol/L HCl), and named E-NAP, U-NAP, Al-NAP, and Ac-NAP. The properties of four NAPs were different. The yields of NAPs were 26.05 % (Ac-NAP) > 20.33 % (Al-NAP) > 17.99 % (U-NAP) > 12.77 % (E-NAP), respectively. The monosaccharide composition of NAPs was composed primarily of mannose, xylose, glucose, glucuronic acid, and galactose. Sequential extraction improved the purity and solubility of NAPs, but decreased the particle size, thermal stability, water retention, and crystallinity. Two polysaccharides, U-NAP and Al-NAP, had a triple helix structure. All the NAPs were pseudoplastic fluids with concentration/frequency-dependent entangled structure. Al-NAP with the highest viscosity exhibited an elastic gel, while Ac-NAP with the lowest viscosity was a viscous gel. The behavior of NAPs differed from that predicted using the Cox-Merz rule, and in particular, E-NAP and U-NAP more significantly deviated from the rule. In this study, four NAPs with different properties were extracted sequentially, which provided a theoretical basis for the down-stream processing with high added-value and utilization of NA and NAP.

All-natural polysaccharide and protein complex nanoparticles from Clitocybe squamulosa as unique Pickering stabilizers for oil-in-water emulsions.

This study aimed to develop novel nanoparticles that can serve as an excellent oil-in-water (O/W) Pickering stabilizer. The polysaccharide-protein complex nanoparticles (PPCNs-20 and PPCNs-40) were prepared at different ultrasonication amplitudes (20 % and 40 %, respectively) from the polysaccharide-protein complexes (PPCs) which were extracted from the residue of Clitocybe squamulose. Compared with PPCs and PPCNs-20, the PPCNs-40 exhibited dispersed blade and rod shape, smaller average size, and larger zeta potential, which indicated significant potential in O/W Pickering emulsion stabilizers. Subsequently, PPCNs-40 stabilized Pickering emulsions were characterized at different concentrations, pHs, and oil phase contents. The average size, micromorphology, rheological properties, and storage stability of the emulsions were improved as the concentration of PPCNs-40, the ratio of the soybean oil phase and pH value increased. Pickering emulsions showed the best stability when the concentration of PPCNs-40 was 3 wt%, and the soybean oil fraction was 30 % under both neutral and alkaline conditions. The emulsions demonstrated shear thinning and gelation behavior. These findings have implications for the use of eco-friendly nanoparticles as stabilizers for Pickering emulsions and provide strategies for increasing the added value of C. squamulosa.

Effect of Different Drying Methods on the Quality of Oudemansiella raphanipes.

In this study, we used fresh Oudemansiella raphanipes as raw materials and pre-treated through hot air drying (HD), infrared radiation drying (ID), and vacuum freeze drying (VD) to investigate the effects of different drying methods on the rehydration rate, appearance quality, microstructure, and volatile flavor components of the dried products, as well as to determine the physicochemical properties and bioactivities of the polysaccharides in the dried O. raphanipes. The results showed that the VD O. raphanipes had the highest rehydration rate and the least shrinkage in appearance, and it better maintained the original color of the gills, but their aroma was not as strong as that of the HD samples. The scanning electron microscopy results indicate that VD maintains a good porous structure in the tissue, while HD and ID exhibit varying degrees of shrinkage and collapse. Seventy-five common volatile substances were detected in the three dried samples, mainly alkanes, alcohols, and esters. The polysaccharides (PS-H, PS-I, and PS-V) extracted from the dried samples of these three species of O. raphanipes had similar infrared spectral features, indicating that their structures are basically consistent. The highest yield was obtained for PS-V, and the polysaccharide content and glucuronic acid content of PS-I were higher than those of the remaining two polysaccharides. In addition, PS-V also showed better antioxidant activity and inhibitory activity against α-glucosidase as well as α-amylase. In conclusion, among the above three drying methods, the quality of O. raphanipes obtained by vacuum freeze drying is the best, and this experiment provides a theoretical basis for the selection of drying methods for O. raphanipes.

Effects of different extraction methods on the physico-chemical characteristics and biological activities of polysaccharides from Clitocybe squamulosa.

This study comparatively evaluated the effects of the commonly used six extraction methods (acidic, alkaline, enzymatic, ultrasonic, high-pressure, and microwave) on the physico-chemical properties, processing characteristics, and biological activities of polysaccharides from Clitocybe squamulosa (CSFPs). The results show that polysaccharides extracted using an enzyme-assisted extraction method has a relatively high extraction yield (4.46 ± 1.62 %) and carbohydrate content (70.79 ± 6.25 %) compared with others. Furthermore, CSFPs were all composed of glucose, galactose, mannose, xylose, and glucosamine hydrochloride. Only ultrasonic-assisted extraction of polysaccharides (CSFP-U) has a triple helix chain conformation. Scanning electron microscopy (SEM) revealed significant differences in the microstructure of polysaccharides prepared using different methods. Besides that, the polysaccharides prepared by alkali extraction (CSFP-B) and high-pressure assisted extraction (CSFP-H) have good water (2.86 ± 0.29 g/g and 3.15 ± 0.29 g/g) and oil (8.13 ± 0.32 g/g and 7.97 ± 0.04 g/g) holding properties. The rheological behavior demonstrated that CSFPs solutions were typical non-Newtonian fluid. Apart from this, the antioxidant capacity (clearing DPPH (IC50 = 0.29) and ABTS free radicals (IC50 = 0.19), total reduction ability (IC50 = 3.02)) of polysaccharides prepared by the microwave-assisted extraction (CSFP-M) method was significantly higher than that of other extraction methods. By contrast, the polysaccharide prepared by acid extraction (CSFP-A) has the optimum binding capacity (bile acid salt (71.30 ± 6.78 %) and cholesterol (57.07 ± 3.26 mg/g)). The antibacterial activity of CSFPs was positively correlated with their concentration. Thus, the research results can provide a theoretical basis for the development and utilization of polysaccharides from C. squamulosa.

The Effects of Naematelia aurantialba on the Pasting and Rheological Properties of Starch and the Research and Development of Soft Candy.

To study the effects of Naematelia aurantialba (NA) on the rheological and gelatinization properties of starch, the processing methods of NA were diversified. In this study, the gelatinization and rheological properties of corn starch (CS) and edible cassava starch (ECS) were investigated by adding NA with different mass fractions. Starch soft candy was prepared using NA, CS, and ECS as the main raw materials. Rheological studies showed that both CS-NA and ECS-NA exhibited elastic modulus (G') > viscosity modulus (G″), implying elastic behavior. G' was such that CS+1%NA > CS+5%NA > CS+3%NA > CS > CS+2%NA > CS+4%NA > ECS+4%NA > ECS+3%NA > ECS+5%NA > ECS+2%NA > ECS+1%NA > ECS. The gelatinization implied showed that after adding NA, the pasting temperature of CS-NA and ECS-NA increased by 1.33 °C and decreased by 2.46 °C, while their breakdown values decreased by 442.35 cP and 866.98 cP, respectively. Through a single-factor test and orthogonal test, the best formula of starch soft candy was as follows: 0.4 f of NA, 10 g of white granulated sugar, a mass ratio of ECS to CS of 20:1 (g:g), 0.12 g of citric acid, 1 g of red date power, and 16 mL of water. The soft candy was stable when stored for two days. This study offers a new direction for the research and development of NA starch foods.

Effects of Freeze-Thaw Cycles on the Structures and Functional Properties of Clitocybe squamulosa Protein Isolates.

Changes in the functional properties and structures of Clitocybe squamulosa protein isolate (CSPI) in the process of freeze-thaw (F-T) cycles were explored. Remarkable alterations and the reduced content of protein ordered structure were revealed through structural analysis of CSPI after F-T treatments. The surface hydrophobicity and free sulfhydryl content of CSPI first increased and then decreased. However, after the F-T treatments, the carbonyl content of CSPI continued to increase. Similarly, the water holding capacity (WHC), oil holding capacity (OHC), and solubility of CSPI all declined as the number of F-T cycles increased. The foaming properties and emulsifying properties of CSPI were significantly improved and reached maximum values after three F-T cycles. CSPI undergoing two F-T cycles showed the highest digestibility, maximum polypeptide content, and highest DPPH and ·OH-radical-scavenging activities. The ·OH-radical-scavenging activities and reducing power of the gastrointestinally digested CSPI had the highest value after one F-T cycle. Therefore, it has been demonstrated that F-T treatments could be a residue-free and cost-effective tool for improving mushroom protein functional properties.

Sparassis latifolia polysaccharides inhibit colon cancer in mice by modulating gut microbiota and metabolism.

Sparassis latifolia polysaccharides (SLPs) can regulate inflammatory cytokines. However, little is known about the regulation mechanism of SLPs on colon cancer. In this study, we investigated the mechanism of SLPs on metabolism in mice with colon cancer. The results showed that SLPs could improve the colon morphology and physiological indices, and inhibit the infiltration of immune cells in colon. Moreover, it could improve metabolism disorder of colon cancer via reducing the levels of TNF-α, IL-6, NF-κB, COX-2 and IL-1ß mRNA or protein, increasing IκB mRNA or protein expression. In addition, it could comprehensively regulate the colon cancer related metabolism by changing the abundance of key intestinal flora and 35 metabolites including phosphatidylcholine, tryptophan and tetrahydrobiopterin. Some biomarkers associated with colon cancer metabolism were related significantly with the abundance of specific intestinal flora. These findings indicate that SLPs can attenuate metabolism disorder of colon cancer by modulating gut microbiota and metabolites.

Effects of in vitro digestion and fecal fermentation on physico-chemical properties and metabolic behavior of polysaccharides from Clitocybe squamulosa.

The aim of this study was to establish a human digestion model in vitro to explore the degradation characteristics of a novel high-purity polysaccharide from Clitocybe squamulosa (CSFP2). The results showed that the content of reducing sugars (CR ) of CSFP2 increased from 0.13 to 0.23 mg/mL, the molecular weight (Mw) of CSFP2 decreased significantly during the saliva-gastrointestinal digestion. The constituent monosaccharides of CSFP2, including galactose, glucose, and mannose, were stable during in vitro digestion, but their molar ratios were changed from 0.023: 0.737: 0.234 to 0.496: 0.478: 0.027. The surface of CSFP2 changes from a rough flaky structure to a scattered flocculent or rod-shaped structure after the gastrointestinal digestion. However, the apparent viscosity of CSFP2 was overall stable during in vitro digestion. Moreover, CSFP2 still maintains its strong antioxidant capacity after saliva-gastrointestinal digestion. The results showed that CSFP2 can be partially decomposed during digestion. Meanwhile, some physico-chemical properties of the fermentation broth containing CSFP2 changed significantly after gut microbiota fermentation. For example, the pH value (from 8.46 to 4.72) decreased significantly (p < 0.05) after 48 h of fermentation. the OD 600 value increased first and then decreased (from 2.00 to 2.68 to 1.32) during 48-h fermentation. In addition, CSFP2 could also increase the amounts of short-chain fatty acids (SCFAs) (from 5.5 to 37.15 mmol/L) during fermentation (in particular, acetic acid, propionic acid, and butyric acid). Furthermore, the relative abundances of Bacteriodes, Bifidobacterium, Catenibacterium, Lachnospiraceae_NK4A136_group, Megasphaera, Prevotella, Megamonas, and Lactobacillus at genus level were markedly increased with the intervention of CSFP2. These results provided a theoretical basis for the further development of functional foods related to CSFP2.

Two Novel Polysaccharides From Clitocybe squamulosa: Their Isolation, Structures, and Bioactivities.

The crude polysaccharides from the fruiting bodies of Clitocybe squamulosa (CSFP) were isolated by hot-water extraction. Two novel polysaccharides, CSFP1-ß 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-ß, 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-ß and CSFP2-α were interwoven by reticular and fibrous structures, respectively. Purified components of both CSFP1-ß 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-ß 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-ß 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-ß, 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.

Effects of different cooking methods on volatile flavor compounds, nutritional constituents, and antioxidant activities of Clitocybe squamulosa.

To explore a scientific and reasonable cooking method for Clitocybe squamulosa, four cooking methods (boiling, steaming, microwaving, and frying) were applied to C. squamulosa, and the effects of different cooking methods on volatile flavor compounds, nutritional constituents, and antioxidant activities in C. squamulosa were systematically investigated. The results showed that 54, 53, 61, 63, and 49 volatile flavor compounds were detected in raw, boiled, steamed, microwaved, and fried samples, respectively. Large differences in volatile flavor compounds between the four cooking and raw samples were determined by using relative odor activity values (ROAV) and principal component analysis (PCA). In addition, steaming and microwaving could protect the nutrients of C. squamulosa, reduce losses during the cooking process and improve the color of cooked products compared to boiling and frying cooking methods. Meanwhile, cooking treatment exerted different effects on the antioxidant activity of C. squamulosa, and the antioxidant activity of C. squamulosa was the highest after microwave cooking treatment. This research can provide a theoretical basis for the cooking, processing and utilization of C. squamulosa and other wild edible fungi.

Ultrasound-Assisted Deep Eutectic Solvents Extraction of Polysaccharides From Morchella importuna: Optimization, Physicochemical Properties, and Bioactivities.

In this study, a high-efficiency and non-pollution extraction procedure, ultrasound-assisted technique with deep eutectic solvents (DESs), was applied for extraction of polysaccharides from Morchella importuna (MIP-D). The results exhibited that the system of DES was: mole ratio between choline chloride and oxalic acid of 2:1, water content of 90% (v/v), and the optimal extraction parameters were as follows: extraction time of 31.2 min, extraction temperature of 62.1°C, and the liquid-solid ratio of 32.5:1 (v/w). Under these extraction parameters, the extraction yield of MIP-D was 4.5 times higher than hot water extraction (HWE) method and had higher carbohydrate (85.27%) and sulfate contents (34.16%). Moreover, high-performance liquid chromatography (HPLC) and Fourier-transform IR (FTIR) spectrum analysis indicated that MIP-D was comprised of glucosamine, galactose, glucose, and mannose, with molar ratios of 0.39:1.88:3.82:3.91, which contained the pyranose ring skeleton. High-performance gel permeation chromatography (HPGPC) analysis revealed that MIP-D showed three fractions with molecular weights of 2.6 × 106, 7.3 × 104, and 3.7 × 103 Da, which were lower than those of polysaccharides extracted by HWE. In-vitro tests proved that MIP-D possessed excellent antioxidant and inhibited α-amylase and α-glucosidase inhibitory activities. Therefore, DESs (choline chloride-oxalic acid) as a high-efficiency and non-pollution solvent alternative can be applied to the separation of bioactive polysaccharides from Morchella importuna (M. importuna).

Purification, Characterization, and Immobilization of a Novel Protease-Resistant α-Galactosidase from Oudemansiella radicata and Its Application in Degradation of Raffinose Family Oligosaccharides from Soymilk.

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.