β-Glucan Assay Kit (Yeast and Mushroom)

A robust and reliable method has been developed for the measurement of β-glucan in mushroom and mycelial products. Total glucan (plus free glucose and glucose from sucrose) was measured using controlled acid hydrolysis with H₂SO₄ and the glucose released specifically was measured using glucose oxidase/peroxidase reagent. α-Glucan (starch/glycogen) plus free glucose and glucose from sucrose were specifically measured after hydrolysis of starch/glycogen to glucose with glucoamylase and sucrose to glucose plus fructose with invertase and the glucose specifically measured with GOPOD reagent. β-Glucan was determined by the difference. Several acid and enzyme-based methods for the hydrolysis of the β-glucan were compared, and the best option was the method using H₂SO₄. For most samples, similar β-glucan values were obtained with both the optimized HCl and H₂SO₄ procedures. However, in the case of certain samples, specifically Ganoderma lucidum and Poria cocus, the H₂SO₄ procedure resulted in significantly higher values. Hydrolysis with 2 N trifluoroacetic acid at 120°C was found to be much less effective than either of the other two acids evaluated. Assays based totally on enzymatic hydrolysis, in general, yielded much lower values than those obtained with the H₂SO₄ procedure.

The oyster mushroom (Pleurotus species) is a popular and widely cultivated edible mushroom that can be found worldwide, including in Malaysia. However, its local production is unable to fulfil the market demand, partly due to the limited availability of rubber wood sawdust (RWS) as the conventional cultivation substrate. Furthermore, the palm oil industry in Malaysia generates large volumes of organic by-products that have caused environmental concerns. Therefore, the potential utilisation of oil palm waste-based substrates in order to develop a substitute RWS for Pleurotus ostreatus mushroom production is evaluated in this study, based on their agronomic performance and nutritional properties. Empty fruit bunches (EFBs), oil palm fronds (OPFs), and oil palm trunks (OPTs) were used to formulate the substrates. The control used was 100% RWS. Generally, 100% EFB showed a better agronomic performance, and mushroom growth was 1.9 times faster compared to the control, with a comparable mushroom yield. The crude protein and beta glucan content of mushrooms grown on oil palm by-product-formulated substrates were significantly higher than those grown using the control. Additionally, the number of fruiting bodies, crude protein, and beta glucan content of the mushrooms were positively correlated with potassium in the substrate. Therefore, 100% EFB could be used as a potential substitute for RWS for the cultivation and production of P. ostreatus.

Fermentation can be a powerful tool for developing new sustainable foods with increased nutritional value and fermented microbial biomass derived from filamentous fungi is a promising example. This study investigates the nutritional profile of edible Aspergillus oryzae biomass produced under submerged fermentation (SmF) using oat flour as a substrate. The fermentation occurred in a 1m³ airlift bioreactor during 48 h at 35°C and the nutritional profile of the produced fungal biomass in terms of amino acids, fatty acids, minerals (Fe, Zn, Cu, Mn), vitamins (E, D₂), and dietary fiber was compared to oat flour as well as pure fungal biomass grown on semi-synthetic medium. The total amount of amino acids increased from 11% per dry weight (dw) in oat flour to 23.5% dw in oat fungal biomass with an improved relative ratio of essential amino acids (0.37 to 0.42). An increase in dietary fibers, minerals (Fe, Zn, Cu), vitamin E, as well as vitamin D₂ were also obtained in the oat fungal biomass compared to oat flour. Moreover, the short chain omega-3 α-linolenic acid (ALA) and omega-6 linoleic acid (LA) values increased from 0.6 to 8.4 and 21.7 to 68.4 (mg/g dry weight sample), respectively, in oat fungal biomass. The results indicate that fungal biomass grown on oat flour could have a potential application in the food industry as a nutritious source for a wide variety of products.

The fruiting bodies of edible mushrooms represent an important source of biologically active polysaccharides. In this study, Lentinula edodes crude polysaccharides (LECP) were extracted in hot water, and their antioxidant and antiradical activities were investigated. The antioxidant activity of LECP was investigated against reactive species such as 1,1’-diphenyl-2-picrylhydrazyl, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, hydroxyl and superoxide anion radicals, reducing power with EC₅₀ values of 0.51, 0.52, 2.19, 3.59 and 1.73 mg/mL, respectively. Likewise, LECP inhibited the lipid peroxidation induced in methyl linoleate through the formation of conjugated diene hydroperoxide and malondialdehyde. The main sugar composition of LECP includes mannose, galactose, glucose, fucose and glucuronic acid. Characterization by Fourier transform infrared spectroscopy and nuclear magnetic resonance determined that LECP was made up of α and β glycosidic bonds with a backbone of α-D-Glc, →6)-β-D-Glcp-(1→, →6)-α-D-Galp-(1→ and β-D-Manp-(1→ residues. The results showed that LECP can scavenge all reactive species tested in a concentration-dependent manner and with a protective effect in the initial and final stages of lipid peroxidation. The natural antioxidant activity of the LECP that was investigated strengthens the high medicinal and nutritional value of this mushroom.

This study aimed to investigate the effects of temperature (100°C–140°C), pressure (0.4-1.0 MPa), and extraction time (20-60 min) on the extraction yield of crude polysaccharides, β-glucan, and total phenolic compounds with pressurized hot water extraction of gray oyster mushrooms (Pleurotus sajor-caju (Fr.) Singer). The extraction conditions were optimized by maximizing all responses, using a central composite design (CCD)-based response surface methodology. Temperature mainly affected the increase in the extracted yield and phenolic content. However, exceeded temperature and extraction time negatively affected the β-glucan yield. The optimal extraction conditions were at 140°C, 0.92 MPa, and 40 min with a corresponding extraction yield of 3.20 ± 0.17 g/100 g, as well as β-glucan and total phenolic contents of 43.84 ± 3.86 mg/100 g and 8.49 ± 0.66 mg gallic acid equivalent/100 g dried extract, respectively. The antioxidant activities of extracts were estimated by in vitro biological assays. The SC₅₀ values of DPPH, NO, and ABTS assays were 1.66 ± 0.02, 1.50 ± 0.19, and 6.94 ± 1.04 mg/mL, respectively. This work demonstrates an alternative approach to valorize over-produced mushrooms.

Inonotus obliquus grows in the Northern Hemisphere on some living broadleaved tree species as a pathogen, causing stem rot. In Estonia, the fungus is well known in the Betula species but can also be found on Alnus. Sterile conks of I. obliquus contain different bioactive compounds, but the quantitative and comparative research of these compounds in conks on different host species is limited. In the current work, I. obliquus was isolated and, evidently, determined from Alnus incana (L.) Moench., Alnus glutinosa (L.) Gaertn., and Betula pendula Roth, and the content of bioactive compounds in conks on these hosts were analysed. All the analysed conks sampled from A. incana and B. pendula contained betulin that varied from 111 to 159 µg/g. A significantly (p < 0.05) higher betulinic acid content was found in conks sampled from A. incana when compared with B. pendula: 474–635 and 20–132 µg/g, respectively. However, the conks from Betula were richer in total polyphenols, flavonols, and glucans. The content of inotodiol was quite similar in the conks from A. incana (7455–8961 µg/g) and B. pendula (7881–9057 µg/g). Also, no significant differences in the lanosterol content were found between the samples from these two tree species. To the best of our knowledge, this study is the first investigation of the chemical composition of I. obliquus parasitizing on Alnus. The results demonstrate that the bioactive compounds are promising in conks of I. obliquus growing not only on Betula but also on the Alnus species. It supports the opportunity to cultivate I. obliquus, also on the Alnus species, thus increasing the economic value of growing this tree species in forestry.

In the last decade, in addition to the study of the nutritional composition of mushrooms, the study of biologically active compounds occupies an important place. However, there is a need to find new and lesser known mushrooms species that have biological activity and potential for application in industrial conditions. The aim of this research is to determine the biological activity of aqueous and ethanolic extracts of Suillus granulatus wild mushroom through the determination of the content of total carbohydrates, total, α and β-glucans, as bioactive compounds, as well as determination of cytotoxic activity. Total carbohydrate content was determined using spectroscopic method, total, α and β-glucans were analysed using specific kits, and MTT test (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was used for cytotoxic activity. IR-ATR (infrared spectroscopy - attenuated total reflection) spectra of mushroom extracts were performed, too. Aqueous extracts had a higher content of total carbohydrates as well as glucan and had better cytotoxic activity against HeLa cells, while ethanolic extract of Suillus granulatus was characterized with better cytotoxic activity against HepG2 cells. Based on IR-ATR, the presence of different types of carbohydrates, glucans, proteins, phenols and flavonoids can be observed in aqueous and ethanolic, which is one of the reasons for the differences in their anticancer activity. The analysed extracts are an excellent basis for their further application in various products in order to obtain functional food with enriched biological value. Thus, using natural dietary supplements can significantly affect the positive changes in the health of consumers.

The study compares the impact of freeze- and spray-drying (FD, SD) microencapsulation methods on the content of β-glucan, total polyphenols (TP), total flavonoids (TF), phenolic acids (PA), and antioxidant activity (AA) in commercially β-glucan powder (Pleurotus ostreatus) using maltodextrin as a carrier. Morphology (scanning electron microscopy- SEM), yield, moisture content (MC), and water activity (a_w) were also evaluated in the samples. Our examinations revealed significant structural differences between powders microencapsulated by the drying methods. As compared to non-encapsulated powder, the SD powder with yield of 44.38 ± 0.55% exhibited more reduced (p < 0.05) values for a_w (0.456 ± 0.001) and MC (8.90 ± 0.44%) than the FD one (yield: 27.97 ± 0.33%; a_w: 0.506 ± 0.002; MC: 11.30 ± 0.28%). In addition, the highest values for β-glucan content (72.39 ± 0.38%), TPC (3.40 ± 0.17 mg GAE/g), and TFC (3.07 ± 0.29 mg QE/g) have been detected in the SD powder. Our results allow for the conclusion that the SD microencapsulation method using maltodextrin seems to be more powerful in terms of the β-glucan powder yield and its contents of β-glucan, TP, and TF as compared to the FD technique.

The consumption of functional foods, such as mushrooms, apparently influences Gestational Diabetes Mellitus (GDM), and brings benefits to maternal-fetal health. Ganoderma lucidum contains a variety of bioactive compounds, such as polysaccharides, proteins and polyphenols that are able to control blood glucose and be used in anti-cancer therapy. We aimed to evaluate the effects of the consumption of Ganoderma lucidum (Gl) on maternal-fetal outcomes in streptozotocin-induced GDM (GDM-STZ). Pregnant rats were exposed to Gl (100 mg/kg/day) before and after the induction of GDM-STZ (single dose 40 mg/kg) on the eighth pregnancy day. Biochemical and oxidative stress parameters, reproductive performance and morphometry of fetuses were assessed. Gl reduced the glycemic response in the oral glucose tolerance test. Moreover, Gl decreased AST and ALT activities. GDM increased lipid peroxidation, which was reverted by Gl. Catalase and glutathione peroxidase activities were decreased in GDM and the administered Gl after the fetus implantation increased catalase activity. Measurements of the fetal head, thorax, craniocaudal and tail showed greater values in fetuses from rats exposed to Gl compared to GDM. Ganoderma lucidum has an encouraging nutritional and medicinal potential against GDM, since it modifies glucose metabolism, reduces lipid peroxidation, and has protective effects in fetuses born from GDM dams.

The incorporation of resistant starch (RS) in food has gained importance to be a good replacement for digestible carbohydrate. This study examined the effect of compositing RS (high-amylose maize starch (HM)) as wheat flour substitute (30%) in Chinese steamed bun (CSB) formulation on postprandial glycemic response in healthy human subject. In this single-blind and cross-over experimental trial, a total of 15 female participants (mean age = 31.5 ± 3.9) were randomly assigned to receive CSB containing 30% HM (HM30) or control CSB (without HM) with their blood glucose were recorded throughout the test. The blood glucose concentrations recorded for HM30 were significantly lower than control CSB at 15 min (6.03 vs. 7.04 mmol/L, p = 0.041), 30 min (6.93 vs. 7.76 mmol/L, p = 0.021), 45 min (6.21 vs. 7.55 mmol/L, p = 0.032), 60 min (5.68 vs. 6.26 mmol/L, p = 0.038), and 90 min (5.08 vs. 5.73 mmol/L, p = 0.022). The 2-h postprandial glucose was significantly lower in HM30 (iAUC = 105.2 mmol x min/L) than the control (186.1 mmol x min/L). The low GI property of HM30 (GI = 39.11 ± 5.6) did not cause sudden rapid increase in blood glucose concentration as observed in medium-GI control CSB (GI = 69.18 ± 9.8). This study suggests that adding 30g of HM decreased the glycemic index of CSB in healthy female adult.

Background: Depression is a severe neuropsychiatric disorder that afects more than 264 million people worldwide. The efcacy of conventional antidepressants are barely adequate and many have side efects. Hericium erinaceus (HE) is a medicinal mushroom that has been reported to have therapeutic potential for treating depression. Methods: Animals subjected to chronic restraint stress were given 4 weeks HE treatment. Animals were then screened for anxiety and depressive-like behaviours. Gene and protein assays, as well as histological analysis were per‑ formed to probe the role of neurogenesis in mediating the therapeutic efect of HE. Temozolomide was administered to validate the neurogenesis-dependent mechanism of HE. Results: The results showed that 4 weeks of HE treatment ameliorated depressive-like behaviours in mice subjected to 14 days of restraint stress. Further molecular assays demonstrated the 4-week HE treatment elevated the expres‑ sion of several neurogenesis-related genes and proteins, including doublecortin, nestin, synaptophysin, brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), phosphorylated extracellular signal-regulated kinase, and phosphorylated cAMP response element-binding protein (pCREB). Increased bromodeoxyuridine-positive cells were also observed in the dentate gyrus of the hippocampus, indicating enhanced neurogenesis. Neurogen‑ esis blocker temozolomide completely abolished the antidepressant-like efects of HE, confrming a neurogenesisdependent mechanism. Moreover, HE induced anti-neuroinfammatory efects through reducing astrocyte activation in the hippocampus, which was also abolished with temozolomide administration. Conclusion: HE exerts antidepressant efects by promoting neurogenesis and reducing neuroinfammation through enhancing the BDNF-TrkB-CREB signalling pathway

This study evaluates the nutritional quality and its biological activity in vivo of a peach palm by-products food ingredient processed via solid-state fermentation by shiitake culinary-medicinal mushroom Lentinula edodes. The group that consumed this diet had higher content of total dietary fiber, digestibility, rate of protein quality and protein efficiency. They also present a late and softer insulinemic peak with an increase in glycemia index, demonstrating amino acids limitation but with feasible matrix as complement proteins. Discrete variation on total cholesterol and triglycerides was observed with a reduction on lipid profile, attributed to its high content of dietary fibre. Lipids from within liver and stools revealed that fermented diet contained the lowest rates of fat in liver and, consequently, highest elimination compared to the other control diets. The serum lipid profile suggests a positive modulation of this diet, and, has good nutritional quality with potential to influence positively the glycaemic and lipid profiles.

Background: The present study extensively aimed to evaluate the underlying mechanism of the immunomodulatory and anti-inflammatory effects of Phellinus linteus mycelium (PLM). Methods: To assess whether PLM influences the production of markers related to inflammation, Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were treated with PLM (50, 100, 200, and 500 μg/mL). Splenocyte, thymus, peritoneal exudate cells (PEC), and peripheral blood mononuclear cells (PBMC) were isolated from the Balb/c mice treated with Korean red ginseng or PLM once a day for 5 weeks. Moreover, all mice except normal mice were stimulated with 10% proteose peptone (PP) treated 3 days before the sacrifice and 2% starch treated 2 days before the sacrifice. Subsequently, the cytotropic substance was evaluated by using flow cytometry analysis and ELISA assay. Results: PLM200 treatment significantly suppressed the production of nitric oxide (NO) and prostaglandin E2 (PGE2) and inhibited the release of proinflammatory cytokines such as interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α dose-dependently in the LPS-stimulated RAW264.7 cells. PLM200 supplementation showed a significant increase in IL-2, IL-12, and interferon (IFN)-γ production and upregulated the ratio of IFN-γ (T-helper type 1, Th1) to IL-4 (T-helper type 2, Th2) in splenocytes. After PLM200 treatment, the significant elevation of CD4⁺CD25⁺, CD4⁺&CD8⁺, and CD4⁺CD69⁺ treatment were detected in thymus. Moreover, CD4⁺ and CD4⁺CD69⁺ in PBMC and CD69⁺ in PEC were also shown in a significant increase. Conclusions: Taken together, these results showed an immunomodulatory effect of PLM about an elevated INF-γ/IL4 ratio, as an index of Th1/Th2, as well as the anti-inflammatory effect in the LPS-stimulated RAW264.7 cells. Therefore, our findings demonstrate that PLM possesses immunostimulatory and anti-inflammatory effects.

Maitake (Grifola frondosa) is a medicinal mushroom known for its peculiar biological activities due to the presence of functional components, including dietary fibers and glucans, that can improve human health through the modulation of the gut microbiota. In this paper, a Maitake ethanol/water extract was prepared and characterized through enzymatic and chemical assays. The prebiotic potential of the extract was evaluated by the growth of some probiotic strains and of a selected probiotic consortium. The results revealed the prebiotic properties due to the stimulation of the growth of the probiotic strains, also in consortium, leading to the production of SCFAs, including lactic, succinic, and valeric acid analyzed via GC-MSD. Then, their beneficials effect were employed in evaluating the vitality of three different healthy and tumoral colorectal cell lines (CCD841, CACO-2, and HT-29) and the viability rescue after co-exposure to different stressor agents and the probiotic consortium secondary metabolites. These metabolites exerted positive effects on colorectal cell lines, in particular in protection from reactive oxygen species.

Probiotic L. acidophilus La-14 cells were co-encapsulated with Ganoderma lingzhi extract to prolong the viability of the cells under simulated gastrointestinal (SGI) condition and to protect the active ingredients of Reishi mushroom during the storage period. Combinations of distinctive reagents (sodium alginate, chitosan, maltose, Hydroxyethyl-cellulose (HEC), hydroxypropyl methylcellulose (HPMC), and calcium lactate) were tested. Optimal double layer Ca-alginate hydrogel beads were fabricated with significantly improved characteristics. The incorporation of maltose significantly decreases the release rate of mushrooms' phenolics, antioxidants, and β-glucan during the storage time. Significant improvement in probiotic cells viability under SGI condition has been found and confirmed by confocal laser microscopy in maltose containing double layer coated calcium alginate beads variants. The encapsulation of newly formulated prebiotic Reishi extract and probiotic L. acidophilus is creating a new potential food application for such medicinal mushrooms and natural products with unpleasant taste upon oral consumption.

The synergic effect using γ-irradiation in combination with hydrogen peroxide was applied for preparation of water-soluble and low molecular weight (Mw) β-glucan from yeast water-insoluble β-glucans. The structural characterization by gel permeation chromatography (GPC), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) suggested that the γ-irradiation did not cause any change in basic structure of β-glucans, except for the reduction in degree of polymerization. The suitable degradation conditions were determined at 10% β-glucans in pH~9% and 1% hydrogen peroxide. The water-soluble oligoβ-glucan with Mw~15 kDa showed a significantly effect on the increase of body weight and survival rate of Penaeus monodon shrimps challenged with Vibrio parahaemolyticus and White spot syndrome virus (WSSV). The oral administration of 15 kDa-oligoβ-glucan with a daily dose of 1000 ppm strongly stimulated immune factors in tested shrimps such as phagocytosis activity (PA), phenoloxidase (PO) and superoxide dismutase (SOD). Furthermore, dietary of 1000 ppm 15 kDa-oligoβ-glucan also significantly up-regulated the gene expressions of lipopolysaccharide and β-1,3-glucan-binding protein (LGLP) and prophenoloxidase (proPO). The relative percent of survivals (RPS) of shrimps fed with 1000 ppm 15 kDa-oligoβ-glucan were 56.2% and 38.1% post the challenge with V. parahaemolyticus and WSSV, respectively. These results demonstrated that the water-soluble oligoβ-glucan with Mw~15 kDa prepared by synergic degradation method showed a very promising potential for application as a natural growth promotor and immunostimulant in shrimp culture.

The purpose of this study was to enhance the immune-enhancing activity of mushroom strains through fermentation to promote food use of leaf extracts of S. quelpaertensis containing β-glucan. We evaluated the immunomodulatory effect of extracts from fermented S. quelpaertensis leaves (SQGL, SQHE, SQPL). S. quelpaertensis leaves fermentation products were prepared by using mushroom mycelia (Ganoderma lucidum, Hericium erinaceum, Phellinus linteus). The content of β-glucan, a major substance in S. quelpaertensis leaves fermentation products, was 3.73 ± 0.50 mg/mL in the extract (SQ) of S. quelpaertensis leaves. The fermented mushrooms, SQGL, were the highest at 5.57 ± 0.86 mg/100 mL, followed by SQHE and SQPL, and the β-glucan content of all of the glucan was >75.3%. To test the immune activity, S. quelpaertensis leaf fermentation products were administered to mice at different doses (60, 160, and 360 mg/kg) for two weeks. Th cell and macrophage populations were found to increase significantly at all three doses compared to the negative control after two weeks. SQGL and SQHE were highest at 160 mg/kg, and SQPL showed the highest Th cell proliferation at 60 mg/kg. In addition, the production of IFN-γ, IL-4, IL-10, and nitric oxide was significantly higher than that of the negative control after two weeks. In particular, an increase was seen at a low concentration of 60 mg/kg. Therefore, the S. quelpaertensis leaf fermentation product can be very useful as a functional ingredient for enhancing immunity.

The effect of heat treatment on dried fruiting bodies of Reishi medicinal mushroom (Ganoderma lingzhi) is investigated. Control and samples treated for 20 min at temperatures of 70, 120, 150 and 180°C were subjected for their free radical scavenging capacity, different glucans and total phenolic content determination. The growth rate of Escherichia coli and Lactobacillus casei supplemented with control and heat-treated samples is also investigated. The roasted mushroom samples at 150°C and 180°C showed the highest level of β-glucan (37.82%) and free radical scavenging capacity on 2,2-diphenyl-1-picrylhidrazyl (DPPH•) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+). The content of total phenolics (TPC) was also influenced by heat treatment and significantly higher TPC values were recorded in samples treated at 120°C and 150°C. The presence of reducing sugars was only detected after heat treatment at 150°C (0.23%) and at 180°C (0.57%). The heat treatments at 120, 150 and 180°C, significantly attenuated the number of colony-forming units (CFU) of pathogenic E. coli, in a linear relationship with an elevated temperature. The supplementation of heat-treated Reishi mushroom at 120°C resulted in the highest growth rate of probiotic L. casei. The obtained results in this study revealed the significant effect of short-term heat treatment by enhancing the antioxidant capacity, β-glucan solubility and prebiotic property of the dried basidiocarp of Reishi mushroom.

Sorghum biscuits were enriched with mushroom powders (Lentinula edodes, Auricularia auricula and Tremella fuciformis) at 5%, 10% and 15% substitution levels. An in vitro gastrointestinal digestion was used to evaluate the effect of this enrichment on the phenolic content and soluble peptide content as well as antioxidant activities of the gastric or intestinal supernatants (bio-accessible fractions), and the remaining portions of phenolic compounds, antioxidants and β-glucan in the undigested residue (non-digestible fraction). The phenolic content of the gastric and intestinal supernatants obtained from digested mushroom-enriched biscuits was found to be higher than that of control biscuit, and the phenolic content was positively correlated to the antioxidant activities in each fraction (p < 0.001). L. edodes and T. fuciformis enrichment increased the soluble protein content (small peptide) of sorghum biscuits after in vitro digestion. All mushroom enrichment increased the total phenolic content and β-glucan content of the undigested residue and they were positively correlated (p < 0.001). The insoluble dietary fibre of biscuits was positively correlated with β-glucan content (p < 0.001) of undigested residue. These findings suggested that enriching food with mushroom derived dietary fibre increases the bioavailability of the non-digestible β-glucan and phenolic compounds.