Novel Catechol O-methyltransferases from Lentinula edodes Catalyze the Generation of Taste-Active Flavonoids.

Due to the increasing demand for natural food ingredients, including taste-active compounds, enzyme-catalyzed conversions of natural substrates, such as flavonoids, are promising tools to align with the principles of Green Chemistry. In this study, a novel O-methyltransferase activity was identified in the mycelium of Lentinula edodes, which was successfully applied to generate the taste-active flavonoids hesperetin, hesperetin dihydrochalcone, homoeriodictyol, and homoeriodictyol dihydrochalcone. Furthermore, the mycelium-mediated OMT activity allowed for the conversion of various catecholic substrates, yielding their respective (iso-)vanilloids, while monohydroxylated compounds were not converted. By means of a bottom-up proteomics approach, three putative O-methyltransferases were identified, and subsequently, synthetic, codon-optimized genes were heterologously expressed in Escherichia coli. The purified enzymes confirmed the biocatalytic O-methylation activity against targeted flavonoids containing catechol motifs.

Peach palm shells (Bactris gasipaes Kunth) bioconversion by Lentinula edodes: Potential as new bioproducts for beef cattle feeding.

This paper aims to develop and assess the in vitro effects on ruminal fermentation and greenhouse gas parameters of new bioproducts for beef cattle diets, carried out by solid-state fermentation of peach palm shells colonized by Lentinula edodes (SSF) and after Shiitake mushroom cultivation in axenic blocks (SMS). In vitro experiments were performed to assess the in vitro gas production, digestibility, and fiber degradation of formulated total diets. Bioproducts presented high ß-glucans (9.44---11.27 %) and protein (10.04---8.35 %) contents, as well as similar digestibility to conventional diets. SMS diet had the lowest methane and carbon dioxide (19.1 and 84.1 mM/g OM) production, and the SSF diet presented lower carbon dioxide production (98.9 mM/g OM) than other diets, whereas methane was similar. This study highlighted a sustainable use of byproducts for beef cattle diets, promising for digestibility, nutritional value, ß-glucans incorporation, and environmental impact mitigation, favoring the circular bioeconomy.

Production of Extracellular Enzymes by Lentinula edodes Strains in Solid-State Fermentation on Lignocellulosic Biomass Sterilized by Physical and Chemical Methods.

Two methods of sterilization of lignocellulosic biomass were performed in this study. Eucalypt waste (EW) supplemented with rice bran (RB) was added in the proportions 80:20 and 90:10 in dry weight. The compositions were sterilized by physical method (autoclaving) and by chemical method (H2O2). The production of extracellular enzymes by Lentinula edodes strains was compared within the two methods. Inactivation of catalase present in RB was achieved with 250 mM H2O2. The use of H2O2, when compared by physical method, favored high production of hydrolytic enzymes such as endoglucanase (1,600 IU/kg), twofold higher, ß-glucosidase (1,000 IU/kg), fivefold higher, xylanase (55,000 IU/kg), threefold higher and ß-xylosidase (225 IU/kg), similar results. Oxidative enzymes, MnP and laccase, were produced within a different profile between strains, with shorter times for laccase (2,200 IU/kg) by SJC in 45 days and MnP (2,000 IU/kg) by CCB-514 in 30 days. High production of extracellular enzymes is achieved by the use of the chemical method of sterilization of lignocellulosic biomass; in addition to no energy consumption, this process is carried out in a shorter execution time when compared to the physical process. The use of H2O2 in sterilization does not produce toxic compounds from the degradation of hemicellulose and cellulose such as furfural and hydroxy-methyl-furfural that cause inhibition of microorganisms and enzymes.

Lentinula edodes Cultured Extract and Rouxiella badensis subsp. acadiensis (Canan SV-53) Intake Alleviates Immune Deregulation and Inflammation by Modulating Signaling Pathways and Epigenetic Mechanisms.

Puberty is a critical developmental period of life characterized by marked physiological changes, including changes in the immune system and gut microbiota development. Exposure to inflammation induced by immune stressors during puberty has been found to stimulate central inflammation and lead to immune disturbance at distant sites from the gut; however, its enduring effects on gut immunity are not well explored. Therefore, in this study, we used a pubertal lipopolysaccharides (LPS)-induced inflammation mouse model to mimic pubertal exposure to inflammation and dysbiosis. We hypothesized that pubertal LPS-induced inflammation may cause long-term dysfunction in gut immunity by enduring dysregulation of inflammatory signaling and epigenetic changes, while prebiotic/probiotic intake may mitigate the gut immune system deregulation later in life. To this end, four-week-old female Balb/c mice were fed prebiotics/probiotics and exposed to LPS in the pubertal window. To better decipher the acute and enduring immunoprotective effects of biotic intake, we addressed the effect of treatment on interleukin (IL)-17 signaling related-cytokines and pathways. In addition, the effect of treatment on gut microbiota and epigenetic alterations, including changes in microRNA (miRNA) expression and DNA methylation, were studied. Our results revealed a significant dysregulation in selected cytokines, proteins, and miRNAs involved in key signaling pathways related to IL-17 production and function, including IL-17A and F, IL-6, IL-1ß, transforming growth factor-ß (TGF-ß), signal transducer and activator of transcription-3 (STAT3), p-STAT3, forkhead box O1 (FOXO1), and miR-145 in the small intestine of adult mice challenged with LPS during puberty. In contrast, dietary interventions mitigated the lasting adverse effects of LPS on gut immune function, partly through epigenetic mechanisms. A DNA methylation analysis demonstrated that enduring changes in gut immunity in adult mice might be linked to differentially methylated genes, including Lpb, Rorc, Runx1, Il17ra, Rac1, Ccl5, and Il10, involved in Th17 cell differentiation and IL-17 production and signaling. In addition, prebiotic administration prevented LPS-induced changes in the gut microbiota in pubertal mice. Together, these results indicate that following a healthy diet rich in prebiotics and probiotics is an optimal strategy for programming immune system function in the critical developmental windows of life and controlling inflammation later in life.

Effect of oxyresveratrol on the quality and membrane lipid metabolism of shiitake mushroom (Lentinus edodes) during storage.

The effect of oxyresveratrol on postharvest quality and membrane lipid metabolism of shiitake mushroom was investigated. The result exhibited that oxyresveratrol retarded browning, maintained firmness and alleviated occurrence of decay of shiitake mushroom. The oxidation and hydrolysis of membrane phospholipids were suppressed by oxyresveratrol treatment, which was associated with reduced LOX and PLD activities and increased SOD and CAT activities. The membrane lipidomics of shiitake mushroom was determined by LC-MS. 385 lipid species and 13 fatty acids in membrane lipids were identified by multiple reaction monitoring method. Compared with control group, the phospholipic acid and lysophospholipid reduced by 29.24% and 21.29% in oxyresveratrol-treated group, respectively, which alleviated hydrolysis of phospholipid. Meanwhile, oxyresveratrol maintained the unsaturation of fatty acids and alleviated oxidation of phospholipid. These results demonstrated that oxyresveratrol could play a dual role of inhibiting the oxidation and hydrolysis of phospholipids to mitigate cellular damage of shiitake mushroom.

Potential Role of Lysine Acetylation and Autophagy in Brown Film Formation and Postripening of Lentinula edodes Mycelium.

Lentinula edodes is one of the most widely cultivated edible mushrooms in the world. When cultivated in sawdust, the surface mycelium of L. edodes needs a long postripening stage wherein it forms a brown film (BF) by secreting and accumulating pigments. BF formation is critical for the high quality and yield of fruiting bodies. Protein lysine acetylation (KAC) is an important post-translational modification that regulates growth and development. Previous studies have shown that deacetylase levels are significantly increased during BF formation in the postripening stage of L. edodes. The aim of this study was to assess the role of protein acetylation during BF formation. To this end, we compared the acetylome of L. edodes mycelia before and after BF formation using anti-acetyl antibody-based label-free quantitative proteomics. We identified 5,613 acetylation sites in 1,991 proteins, and quantitative information was available for 4,848 of these sites in 1,815 proteins. Comparative acetylome analysis showed that the modification of 699 sites increased and that of 562 sites decreased during BF formation. Bioinformatics analysis of the differentially acetylated proteins showed significant enrichment in the tricarboxylic acid (TCA) cycle and proteasome pathways. Furthermore, functional assays showed that BF formation is associated with significant changes in the activities of proteasome, citrate synthase, and isocitrate dehydrogenase. Consistent with this hypothesis, the lysine deacetylase inhibitor trichostatin (TSA) delayed autophagy and BF formation in L. edodes. Taken together, KAC and autophagy play important roles in the mycelial BF formation and postripening stage of L. edodes. IMPORTANCE Mycelial BF formation and postripening of L. edodes affects the quality and quantity of its edible fruiting bodies. In this study, we explored the role of protein KAC in this biological process, with the aim of optimizing the cultivation and yield of L. edodes.

Effects of continuous enzymolysis on the umami characteristics of Lentinula edodes and the flavor formation mechanism of umami peptides.

The purpose of this study was to explore the effect of continuous enzymolysis on the umami characteristics of Lentinula edodes and illuminate the umami mechanism of peptides. The results indicated that the continuous enzymolysis extracts (LFTE) of L.edodes had higher umami intensity and palatability than the water extracts (LWE). 1H NMR and LC-MS/MS were used to evaluate taste metabolites and peptide profiles. Among the identified peptides, LPGVAE, LDELEK, DVELSK, LPDEAR, and TTLPDK with high umami scores which threshold in the range of 0.091-0.371 mmol/L were screened by iUmami-SCM and BIOPEP-UWM, and further verified by sensory evaluation. The results of molecular docking suggested that Ser148, Asn150, Ser276, Ser278 of T1R1 and Asn68, Val277, Ala302, Ser306 of T1R3 played a key role in the umami peptides docking. The study revealed continuous enzymolysis of L.edodes could obtain more umami substances and umami peptides, which laid a foundation for researching flavor substances and developing flavor products from L.edodes.

Proteomic characterization of Shiitake (Lentinula edodes) post-harvest fruit bodies grown on hardwood logs and isolation of an antibacterial serine protease inhibitor.

Lentinula edodes (Shiitake) is one of the most heavily cultivated mushrooms in the world with proven antioxidant and antibacterial properties, among others. Evidence indicates that the choice of mushroom cultivation technique strongly influences the production of bioactive compounds, but to date the nature of many of these compounds has not been fully established. This work focuses on the proteomic characterization of L. edodes to highlight the main active processes two days after harvest and elucidates the proteins involved in the known antioxidant and antibacterial proprieties of Shiitake fruit bodies cultivated on oak logs. A label-free approach allowed us to identify a total of 2702 proteins which were mainly involved in carbohydrate and protein metabolism, cell growth and replication, indicating that several developmental processes remain active in fruit bodies post-harvest. Proteins with antioxidant activities were identified, indicating the contribution of proteins to the antioxidant properties of L. edodes extracts. Antibacterial assays also reveal the activity of a serine protease inhibitor that strongly accumulates in the post-harvest fruit body grown on oak logs. Overall, this study contributes to the understanding of the impact of the log cultivation method on the production of Shiitake mushrooms richest in high-value bioactive compounds.

Morphological and transcriptomic analysis revealing morphological variations and genetic clues in one Lentinula edodes abnormal browning strain.

Strain abnormal browning is a common problem during cultivation of Lentinula edodes. In this study, the L. edodes strain mycelia isolated from Le-WB and cultured on MYG (Le-WP) isolated from an abnormal browning bag was compared with its normal control mycelia isolated from Le-BB and cultured on MYG (Le-BP). The aerial hyphae of Le-WP were white, and the hyphal growth was significantly reduced. Morphological observation of Le-WP under scanning electron microscope (SEM) and transmission electron microscopy (TEM) revealed abnormal organelle structures. Through transcriptomic analysis, more differentially expressed genes (DEGs) were expressed in the metabolic process and catalytic activity in Le-WP than Le-BP. Two Kyoto encyclopedia of genes and genomes (KEGG) pathways named pentose and glucorunate interconversions, and starch and sucrose metabolism were found to be enriched in Le-WP. The gene expression profiles involved in these two pathways were further analyzed and 12 key genes were selected to be verified by quantitative real-time PCR (qRT-PCR), and the results showed that most of these genes were upregulated in Le-WP. Additionally, the content of 1,3-beta-glucan in Le-WP was also significantly higher than in other samples. This research suggests that abnormal strains may be related to the abnormal synthesis of 1,3-beta-glucan, and it needs further research. This research exhibits possible morphological and genetic clues of Le-WP and lays the foundation for understanding the degeneration of L. edodes strains.

Edible algae (Ecklonia cava) bioprocessed with mycelia of shiitake (Lentinula edodes) mushrooms in liquid culture and its isolated fractions protect mice against allergic asthma.

BACKGROUND: Ecklonia cava is an edible marine brown alga harvested from the ocean that is widely consumed in Asian countries as a health-promoting medicinal food The objective of the present study is to evaluate the anti-asthma mechanism of a new functional food produced by bioprocessing edible algae Ecklonia cava and shiitake Lentinula edodes mushroom mycelia and isolated fractions. METHODS: We used as series of methods, including high performance liquid chromatography, gas chromatography, cell assays, and an in vivo mouse assay to evaluate the asthma-inhibitory effect of Ecklonia cava bioprocessed (fermented) with Lentinula edodes shiitake mushroom mycelium and its isolated fractions in mast cells and in orally fed mice. RESULTS: The treatments inhibited the degranulation of RBL-2H3 cells and immunoglobulin E (IgE) production, suggesting anti-asthma effects in vitro. The in vitro anti-asthma effects in cells were confirmed in mice following the induction of asthma by alumina and chicken egg ovalbumin (OVA). Oral administration of the bioprocessed Ecklonia cava and purified fractions suppressed the induction of asthma and was accompanied by the inhibition of inflammation- and immune-related substances, including eotaxin; thymic stromal lymphopoietin (TSLP); OVA-specific IgE; leukotriene C4 (LTC4); prostaglandin D2 (PGD2); and vascular cell adhesion molecule-1 (VCAM-1) in bronchoalveolar lavage fluid (BALF) and other fluids and organs. Th2 cytokines were reduced and Th1 cytokines were restored in serum, suggesting the asthma-induced inhibitory effect is regulated by the balance of the Th1/Th2 immune response. Serum levels of IL-10, a regulatory T cell (Treg) cytokine, were increased, further favoring reduced inflammation. Histology of lung tissues revealed that the treatment also reversed the thickening of the airway wall and the contraction and infiltration of bronchial and blood vessels and perialveolar inflammatory cells. The bioprocessed Ecklonia cava/mushroom mycelia new functional food showed the highest inhibition as compared with commercial algae and the fractions isolated from the bioprocessed product. CONCLUSIONS: The in vitro cell and in vivo mouse assays demonstrate the potential value of the new bioprocessed formulation as an anti-inflammatory and anti-allergic combination of natural compounds against allergic asthma and might also ameliorate allergic manifestations of foods, drugs, and viral infections.

LENTINULA EDODES POLYSACCHARIDES SUPPRESSED PRO-INFLAMMATORY CYTOKINES EXPRESSION AND COLITIS IN MICE.

BACKGROUND: Polysaccharides from edible mushrooms possess immunomodulatory, anti-inflammatory, and anti-tumor activities. Recent studies indicated that necroptosis plays a role in the pathogenesis of inflammatory diseases and mediates increased expression of inflammatory cytokines. OBJECTIVE: Therefore, it is imperative to determine the impact of polysaccharide extract from Lentinula edodes (L. edodes) on inflammatory cytokines in experimental model of colitis in mice. METHODS: Female C57BL/6 mice divided into three or four mice per group were used for this study. Polysaccharide sample was orally administered to mice prior to (7 days) and during colitis induction with 2.5% dextran sodium sulfate (7 days), followed by additional 3 days of administration. Changes in body weight and colon length were used as markers for colitis, and pro-inflammatory cytokines and tumor necrosis factor receptor 1 (TNFR1) expressions, as well as necroptosis were analyzed in the colon of colitis mice. Data obtained were analysed by Tukey-Kramer and two-tailed standard t tests. RESULTS: The results indicated that the polysaccharide sample suppressed colitis in mice using effects on the body weight and colon length as markers. Also, it was demonstrated that necrostatin-1, a specific inhibitor of necroptosis, suppressed the expression of interleukin (IL)-8, a pro-inflammatory chemokine, in Caco-2 cells induced necroptosis induced by zVAD and TNF-α, an indication that necroptosis may be involved in the expression of pro-inflammatory cytokines. Moreover, the polysaccharide sample suppressed the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, IL-6, IL-1ß, and interferon (IFN)-γ in the colon of mice. CONCLUSION: These results suggested that the suppressive effects of the polysaccharide sample on inflammatory cytokines expression may contribute to its anti-colitis effect, and so may serve as a potent therapeutic agent against inflammatory bowel disease.

Antibacterial activity and antibacterial mechanism of ethanol extracts of Lentinula edodes (Shiitake) and Agaricus bisporus (button mushroom).

The aim of the present work was the comparison of antimicrobial activity, mechanism and components of the ethanol extract (EE) from Lentinula edodes Berk (Pegler) and Agaricus bisporus Sing (Lange). The main component of EE of A. bisporus was dianhydromannitol (20.1%), while isosorbide/dianhydromannitol (21.8%) was detected at a high rate in L. edodes ethanol extract by GC-MS . The common phenolic acids were determined as chlorogenic acid, syringic acid, rutin, p-coumaric acid, ferulic acid, 2-hydroxy cinnamic acid, protocatechuic acid, abscisic acid, and trans-cinnamic acid in both mushroom extract by HPLC-MWD . The MICs (minimum inhibitory concentration) of L. edodes EE on Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis and Acinetobacter baumannii were between 5.1 mg ml-1 and 6.01 mg ml-1, while MICs of A. bisporus EE on the pathogens were between 5.8 mg ml-1 and 9.54 mg ml-1. The highest DRA decrease was in E. faecalis (69.1%) for L. edodes and S. aureus (71.0%) for A. bisporus in the 20th minute. As a result, L. edodes and A. bisporus have a similar antibacterial effect on the pathogens, and this inhibition effect caused DNA, protein leakage and destruction of permeability of bacterial cell membrane by bioactive molecules in mushroom extract.

Spent mushroom substrates for ethanol production - Effect of chemical and structural factors on enzymatic saccharification and ethanolic fermentation of Lentinula edodes-pretreated hardwood.

Spent mushroom substrates (SMS) from cultivation of shiitake (Lentinula edodes) on three hardwood species were investigated regarding their potential for cellulose saccharification and for ethanolic fermentation of the produced hydrolysates. High glucan digestibility was achieved during enzymatic saccharification of the SMSs, which was related to the low mass fractions of lignin and xylan, and it was neither affected by the relative content of lignin guaiacyl units nor the substrate crystallinity. The high nitrogen content in SMS hydrolysates, which was a consequence of the fungal pretreatment, was positive for the fermentation, and it ensured ethanol yields corresponding to 84-87% of the theoretical value in fermentations without nutrient supplementation. Phenolic compounds and acetic acid were detected in the SMS hydrolysates, but due to their low concentrations, the inhibitory effect was limited. The solid leftovers resulting from SMS hydrolysis and the fermentation residues were quantified and characterized for further valorisation.

Proteins and polysaccharides from vegetative mycelium of medicinal basidiomycete Lentinus edodes display cytotoxicity towards human and animal cancer cell lines.

Detection and study of biologically active compounds seems a promising area of research in cancer diagnostics and therapies. The glycoprotein and polysaccharide fractions showing high cytotoxicity towards several human and animal cancer cell lines: A549, Hep-2, HeLa, С6 and SPEV-2 were isolated from basidiomycete Lentinus edodes vegetative mycelium and fruiting body and further characterized. It was found that water-soluble glycoprotein fractions caused the most significant, 70-100% inhibition of metabolic activity of SPЕV-2, А549 and С6 cell lines. The effective concentrations of glycoprotein fractions reducing the viability of cancer cell lines were determined. The protein and subunit composition of fractions was studied; the highly active galactose-specific lectins were found to be present in these fractions. Comparative analysis of transcriptomes of L. edodes vegetative mycelium, fruiting body and primordium revealed the presence of carbohydrate-binding glycoproteins (lectins) specific for each stage of basidiomycete morphogenesis. Histological examination revealed some morphological indicators of immune system activation and the absence of toxic effect on gastro-intestinal mucosa of animals at peroral administration of fungal glycoprotein fractions. Fungal protein and, in particular, lectin preparations derived from L. еdodes vegetative mycelium might be considered as novel prospective tools in cancer diagnostics and therapies.

Effects of fermentation with Lactobacillus fermentum 21828 on the nutritional characteristics and antioxidant activity of Lentinus edodes liquid.

BACKGROUND: Few studies to date have evaluated the use of Lactobacillus and Bifidobacterium in edible fungus fermentation. To obtain a fermented Lentinus edodes liquid product with good taste and effects, a strain with good fermentation performance from nine strains tested was selected, and the physicochemical properties and antioxidant capacity of the resulting product were evaluated. RESULTS: Lactobacillus fermentum 21828 exhibited adhesion, tolerance to low pH and bile salts, and good fermentation performance. The number of viable bacteria was 1.05 × 108 CFU mL-1 , and the extraction rate of crude polysaccharide from L. edodes was 2.79% after fermentation. The effects of fermentation on the contents and composition of nutrients in L. edodes liquid were marked, with changes in total soluble protein, total soluble sugar, total acid, and total phenol levels. The 2,2-diphenyl-1-picrylhydrazyl radical-scavenging rate in the fermentation liquid was 93.01%, which was significantly higher than that in non-fermented liquid (80.33%). Furthermore, analysis of volatile and 5'-nucleotide contents showed that fermentation altered the flavor of the product, whereas sensory evaluation showed that the fermented product was preferred. CONCLUSION: Our study demonstrated that the fermented L. edodes liquid exhibited better nutritional and functional properties, as well as sensory characteristics, compared with unfermented liquid. © 2021 Society of Chemical Industry.

Selenium-Containing Exopolysaccharides Isolated from the Culture Medium of Lentinula edodes: Structure and Biological Activity.

In continuation of our research on the influence of selenium incorporation on the biosynthesis, structure, and immunomodulatory and antioxidant activities of polysaccharides of fungal origin, we have isolated from a post-culture medium of Lentinula edodes a selenium (Se)-containing exopolysaccharide fraction composed mainly of a highly branched 1-6-α-mannoprotein of molecular weight 4.5 × 106 Da, with 15% protein component. The structure of this fraction resembled mannoproteins isolated from yeast and other mushroom cultures, but it was characterized by a significantly higher molecular weight. X-ray absorption fine structure spectral analysis in the near edge region (XANES) suggested that selenium in the Se-exopolysaccharide structure was present mainly at the IV oxidation state. The simulation analysis in the EXAFS region suggested the presence of two oxygen atoms in the region surrounding the selenium. On the grounds of our previous studies, we hypothesized that selenium-enriched exopolysaccharides would possess higher biological activity than the non-Se-enriched reference fraction. To perform structure-activity studies, we conducted the same tests of biological activity as for previously obtained mycelial Se-polyglucans. The Se-enriched exopolysaccharide fraction significantly enhanced cell viability when incubated with normal (human umbilical vein endothelial cells (HUVEC)) cells (but this effect was absent for malignant human cervical HeLa cells) and this fraction also protected the cells from oxidative stress conditions. The results of tests on the proliferation of human peripheral blood mononuclear cells suggested a selective immunosuppressive activity, like previously tested Se-polyglucans isolated from L. edodes mycelium. The Se-exopolysaccharide fraction, in concentrations of 10-100 µg/mL, inhibited human T lymphocyte proliferation induced by mitogens, without significant effects on B lymphocytes. As with previously obtained Se-polyglucans, in the currently tested Se-polymannans, the selenium content increased the biological activity. However, the activity of selenium exopolysaccharides in all tests was significantly lower than that of previously tested mycelial isolates, most likely due to a different mode of selenium binding and its higher degree of oxidation.

Removal of Aflatoxin B1 by Edible Mushroom-Forming Fungi and Its Mechanism.

Aflatoxins (AFs) are biologically active toxic metabolites, which are produced by certain toxigenic Aspergillus sp. on agricultural crops. In this study, five edible mushroom-forming fungi were analyzed using high-performance liquid chromatography fluorescence detector (HPLC-FLD) for their ability to remove aflatoxin B1 (AFB1), one of the most potent naturally occurring carcinogens known. Bjerkandera adusta and Auricularia auricular-judae showed the most significant AFB1 removal activities (96.3% and 100%, respectively) among five strains after 14-day incubation. The cell lysate from B. adusta exhibited higher AFB1 removal activity (35%) than the cell-free supernatant (13%) after 1-day incubation and the highest removal activity (80%) after 5-day incubation at 40 °C. In addition, AFB1 analyses using whole cells, cell lysates, and cell debris from B. adusta showed that cell debris had the highest AFB1 removal activity at 5th day (95%). Moreover, exopolysaccharides from B. adusta showed an increasing trend (24-48%) similar to whole cells and cell lysates after 5- day incubation. Our results strongly suggest that AFB1 removal activity by whole cells was mainly due to AFB1 binding onto cell debris during early incubation and partly due to binding onto cell lysates along with exopolysaccharides after saturation of AFB1 binding process onto cell wall components.

Identification of the Primary Structure of Selenium-Containing Polysaccharides Selectively Inhibiting T-Cell Proliferation.

We previously described the biosynthesis, isolation, and immunosuppressive activity of the selenium-containing polysaccharide fraction isolated from the mycelial culture of Lentinula edodes. Structural studies have shown that the fraction was a protein-containing mixture of high molar mass polysaccharides α- and ß-glucans. However, which of the components of the complex fraction is responsible for the immunosuppressive activity non-typical for polysaccharides of fungal origin has not been explained. In the current study, we defined four-polysaccharide components of the Se-containing polysaccharide fraction determined their primary structure and examined the effect on T- and B-cell proliferation. The isolated Se-polysaccharides, α-1,4-glucan (Mw 2.25 × 106 g/mol), unbranched ß-1,6-d-glucan, unbranched ß-1,3-d-glucan and ß-1,3-branched ß-1,6-d-glucan (Mw 1.10 × 105 g/mol), are not typical as components of the cell wall of L. edodes. All are biologically active, but the inhibitory effect of the isolated polysaccharides on lymphocyte proliferation was weaker, though more selective than that of the crude fraction.

Relationship between flavor and energy status in shiitake mushroom (Lentinula edodes) harvested at different developmental stages.

To understand the relationship between flavor and energy, the flavor, energy, and enzyme activity related to energy metabolism in shiitake mushrooms harvested at different developmental stages were investigated. The results indicated that the adenosine triphosphate level increased significantly in developing mushrooms and was strongly correlated with the fresh weight. The levels of equivalent umami concentration (EUC), total aroma compounds, energy charge, adenosine triphosphatase, cytochrome c oxidase, and succinic dehydrogenase varied with maturity. In addition, a strong correlation was observed between aroma compounds, EUC, and energy status (p < 0.05). Our results suggest that the unique flavor of developing shiitake mushroom is closely related to energy. The findings may provide a new strategy to improve the flavor of mushrooms by regulating their energy levels. PRACTICAL APPLICATION: The unique flavor of shiitake mushroom, which has a significant impact on consumer preferences, is one of its key characteristics. This research paper provides a theoretical foundation for determining the optimal harvest period for shiitake mushrooms with high quality and a new strategy to improve the flavor of mushrooms by regulating their energy levels.

Proteome Analysis and In Vitro Antiviral, Anticancer and Antioxidant Capacities of the Aqueous Extracts of Lentinula edodes and Pleurotus ostreatus Edible Mushrooms.

In this study, we examined aqueous extracts of the edible mushrooms Pleurotus ostreatus (oyster mushroom) and Lentinula edodes (shiitake mushroom). Proteome analysis was conducted using LC-Triple TOF-MS and showed the expression of 753 proteins by Pleurotus ostreatus, and 432 proteins by Lentinula edodes. Bioactive peptides: Rab GDP dissociation inhibitor, superoxide dismutase, thioredoxin reductase, serine proteinase and lectin, were identified in both mushrooms. The extracts also included promising bioactive compounds including phenolics, flavonoids, vitamins and amino acids. The extracts showed promising antiviral activities, with a selectivity index (SI) of 4.5 for Pleurotus ostreatus against adenovirus (Ad7), and a slight activity for Lentinula edodes against herpes simplex-II (HSV-2). The extracts were not cytotoxic to normal human peripheral blood mononuclear cells (PBMCs). On the contrary, they showed moderate cytotoxicity against various cancer cell lines. Additionally, antioxidant activity was assessed using DPPH radical scavenging, ABTS radical cation scavenging and ORAC assays. The two extracts showed potential antioxidant activities, with the maximum activity seen for Pleurotus ostreatus (IC50 µg/mL) = 39.46 ± 1.27 for DPPH; 11.22 ± 1.81 for ABTS; and 21.40 ± 2.20 for ORAC assays. This study encourages the use of these mushrooms in medicine in the light of their low cytotoxicity on normal PBMCs vis à vis their antiviral, antitumor and antioxidant capabilities.