Anti-Inflammatory Sesquiterpenes from Fruiting Bodies of Schizophyllum commune.

Schizophyllum commune, a fleshy fungus, is an important medicinal and food-homologous mushroom in China. In this work, eight undescribed sesquiterpenes schizomycins A-H (1-8) and one new meroterpenoid schizomycin I (9) together with three known analogues (10-12) were isolated from fruiting bodies of S. commune. Their planar structures were established by extensive spectroscopic and mass spectrometric data. The absolute configurations of compounds 1, 2, and 4 were determined by single crystal X-ray diffraction, and compounds 3 and 5-9 were confirmed by electronic circular dichroism calculations. Anti-inflammatory activities of all isolated compounds were evaluated for their inhibitory effects on IL-6 and IL-1ß production in RAW 264.7 cells. Among them, compound 7 exhibited significant IL-6 inhibitory activity with an IC50 value of 3.6 µM. The results of molecular docking showed that compound 7 interacts with amino acid residues (Gly117, Lys118, Asp120, Thr166, and Try168) of the IL-6 receptor protein through hydrogen bonding.

Antifungal Activity of Medicinal Mushrooms and Optimization of Submerged Culture Conditions for Schizophyllum commune (Agaricomycetes).

The main goal of the present study was the exploration of the antifungal properties of Agaricomycetes mushrooms. Among twenty-three tested mushrooms against A. niger, B. cinerea, F. oxysporum, and G. bidwellii, Schizophyllum commune demonstrated highest inhibition rates and showed 35.7%, 6.5%, 50.4%, and 66.0% of growth inhibition, respectively. To reveal culture conditions enhancing the antifungal potential of Sch. commune, several carbon (lignocellulosic substrates among them) and nitrogen sources and their optimal concentrations were investigated. Presence of 6% mandarin juice production waste (MJPW) and 6% of peptone in nutrient medium promoted antifungal activity of selected mushroom. It was determined that, extracts obtained in the presence of MJPW effectively inhibited the grow of pathogenic fungi. Moreover, the content of phenolic compounds in the extracts obtained from Sch. commune grown on MJPW was several times higher (0.87 ± 0.05 GAE/g to 2.38 ± 0.08 GAE/g) than the extracts obtained from the mushroom grown on the synthetic (glycerol contained) nutrient medium (0.21 ± 0.03 GAE/g to 0.88 ± 0.05 GAE/g). Flavonoid contents in the extracts from Sch. commune varied from 0.58 ± 0.03 to 27.2 ± 0.8 mg QE/g. Identification of phenolic compounds composition in water and ethanol extracts were provided by mass spectrometry analysis. Extracts demonstrate considerable free radical scavenging activities and the IC50 values were generally low for the extracts, ranging from 1.9 mg/ml to 6.7 mg/ml. All the samples displayed a positive correlation between their concentration (0.05-15.0 mg/ml) and DPPH radical scavenging activity. This investigation revealed that Sch. commune mushroom has great potential to be used as a source of antifungal and antioxidant substances.

Schizophyllum commune Reduces Expression of the SARS-CoV-2 Receptors ACE2 and TMPRSS2.

The current global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of COVID-19 has infected hundreds of millions of people, killed millions, and continues to pose a threat. It has become one of the largest epidemics in human history, causing enormous damage to people's lives and economies in the whole world. However, there are still many uncertainties and continued attention to the impact of SARS-CoV-2 on human health. The entry of SARS-CoV-2 into host cells is facilitated by the binding of the spike protein on the virus surface to the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Furthermore, transmembrane protease serine 2 (TMPRSS2) is a host surface protease that cleaves and proteolytically activates its S protein, which is necessary for viral infection. Thus, SARS-CoV-2 uses the ACE2 receptor for cell entry and initiates the S protein using the protease TMPRSS2. Schizophyllum commune (SC) is one of the most widely distributed fungi, often found on the rotten wood of trees that has been found to have various health benefits, including anticancer, antimicrobial activity, antiparasitic, and immunomodulatory function. In this article, SC significantly diminished the expression ACE2 and TMPRSS2 protein in vitro and in vivo without cell damage. In addition, adenosine from SC was also proven in this experiment to reduce the ACE2 and TMPRSS2 expression. Thus, our findings suggest that SC and adenosine exhibit potential for the repression of SARS-CoV-2 infection via the ACE2 and TMPRSS2 axis.

Evidence for the Presence of Hyphae and Fruiting Body Calcium Oxalate Crystallites in the Split Gill Medicinal Mushroom Schizophyllum commune (Agaricomycetes).

Biomineralization is the phenomenon by which organisms form crystals. Studies have shown that many fungi can biomineralize, producing calcium oxalate crystals on their hyphae and fruiting body. Schizophyllum commune is a medicinal and edible fungus found worldwide, however, studies on biomineralization in this fungus are limited. Here, samples of Sch. commune fruiting bodies were collected from three different provinces in China and hyphal cells were cultured. Using light microscopy, FE-SEM, and EDAX, we identified crystals on the fruiting body and mycelium of each strain and analyzed their morphological characteristics and ion content. These data demonstrate that biomineralization occurs in Sch. commune in nature as well as during subsequent in vitro culture.

Antibacterial activities of the chemical constituents of Schizophyllum commune MST7-3 collected from coal area.

Two fusidane-type active compounds (6 and 7) and five new ones (1-5), along with other nine known compounds (8-16) were isolated from the metabolites of Schizophyllum commune MST7-3. Their structures were elucidated on the basis of spectroscopic analysis. The absolute configurations of compounds 2 and 3 were established by Mosher's method and optical rotation. Compounds 6 and 7 showed significant antibacterial activities against Stenotrophomonas maltophilia with MIC values of 4 µg/mL and 16 µg/mL, respectively.

Schizophyllum commune ß-glucan: Effect on interleukin-10 expression induced by lipopolysaccharide from periodontopathic bacteria.

ß-glucans are potent immunomodulators, with effects on innate and adaptive immune responses via dectin-1 as the main receptor. In this study, we investigated the biological effect of ß-glucan from Schizophyllum commune, called Schizophyllan (SPG) on Interleukin-10 (IL-10) expression induced by a lipopolysaccharide (LPS) from Aggregatibacter actinomycetemcomitans in murine macrophages (J774.1). SPG and dectin-1 interaction up-regulates LPS-induced IL-10 expression. The regulative effect of SPG on IL-10 expression is dependent on prolongation of nuclear translocation activity of nuclear factor-kappa B (NF-κBα) pathway induced by LPS. We also found that LPS-induced phosphorylation of mitogen- and stress-activated protein kinase 1 (MSK1) and cAMP-responsive-element-binding protein (CREB), followed by up-regulation of IL-10, was stimulated by SPG priming via activation of the spleen tyrosine kinase (Syk). Our data indicate that SPG augments the anti-inflammatory response in murine macrophages which can be useful to create an intervention for periodontal disease treatment.

Screening of Georgian Medicinal Mushrooms for Their Antibacterial Activity and Optimization of Cultivation Conditions for the Split Gill Medicinal Mushroom, Schizophyllum commune BCC64 (Agaricomycetes).

The present study aimed to evaluate the antibacterial activity (ABA) of new mushroom strains collected from the mountain and plain forests of Georgia and belonging to different taxonomic groups. Of 30 Basidiomycetes strains tested on agar plates, Schizophyllum commune BCC64 exhibited the highest inhibitory activity against Escherichia coli and Staphylococcus aureus by the diameter of inhibition zones (17 ± 1 mm and 19 ± 1 mm, respectively). Moreover, this mushroom showed strong activity against Staphylococcus enteritidis (11 mm), Pseudomonas aeruginosa (19 mm), and Salmonella epidermitidis (12 mm). In the submerged cultivation in synthetic medium, xylose and glucose ensured the highest ABA toward S. aureus (70% inhibition in microplate rider tests) and E. coli (60%), respectively. Among lignocellulosic materials tested in the submerged and solid-state fermentation, mandarin marc was found to be an excellent growth substrate for ABA accumulation by Sch. commune 64. Of six nitrogen sources, KNO3 favored the mushroom ABA increase against both bacteria. The suitability of the developed nutrient medium has been proven in 7 L fermenter. After fermentation, ethyl acetate extract obtained from culture liquid and ethanol extract obtained from mycelial biomass of Sch. commune 64 showed the best minimum inhibitory concentration (MIC) against E. coli (0.5 and 2.5 mg/mL, respectively) and S. aureus (1 mg/mL for both extracts).

Chemical features and antioxidant profile by Schizophyllum commune produced on different agroindustrial wastes and byproducts of biodiesel production.

Schizophyllum commune VE_07 was produced in different culture media containing pine sawdust (PS), grape residue (GR), cotton cake (CC) and jatropha seed cake (JC). The content of phenolics and antioxidant activity were determined for the substrates and mushrooms produced. The content of ß-glucans and the composition of S. commune were also evaluated. The medium formulated with 94% grape residue enabled the highest values of yield, biological efficiency, and productivity. Mushrooms grown in this condition showed the highest value (13.14%) of ß-glucans. The contents of proteins and dietary fibre were 16.59% and 59.61%, respectively. Mushrooms grown in cotton cake showed the highest phenolic content (291.51 ± 1.83 mg GAE/ 100 g mushroom) and antioxidant activity (58.15 ± 0.86 DPPH % scavenging). The results obtained indicate that substrate composition affected the production of S. commune and its chemical composition.

Schizophyllum commune induced oxidative stress and immunosuppressive activity in Spodoptera litura.

BACKGROUND: In the last few decades, considerable attention has been paid to fungal endophytes as biocontrol agents, however little is known about their mode of action. This study aimed to investigate the toxic effects of an endophytic fungus Schizophyllum commune by analyzing activities of antioxidant and detoxifying enzymes as well as morphology of haemocytes using Spodoptera litura as a model. RESULTS: Ethyl acetate extract of S. commune was fed to the larvae of S. litura using the artificial diet having 276.54 µg/ml (LC50 of fungus) concentration for different time durations. Exposed groups revealed significant (p ≤ 0.05) increase in the activities of various enzymes viz. Catalase, Ascorbate peroxidase, Superoxide dismutase, Glutathione-S-Transferase. Furthermore, haemocytes showed various deformities like breakage in the cell membrane, cytoplasmic leakage and appearance of strumae in the treated larvae. A drastic reduction in the percentage of normal haemocytes was recorded in the treated groups with respect to control. CONCLUSION: The study provides important information regarding the oxidative stress causing and immunosuppressant potential of S. commune against S. litura and its considerable potential for incorporation in pest management programs.

Chitosan-glucan complex hollow fibers reinforced collagen wound dressing embedded with aloe vera. II. Multifunctional properties to promote cutaneous wound healing.

This study presents an innovative multifunctional system in fabricating new functional wound dressing (FWD) products that could be used for skin regeneration, especially in cases of infected chronic wounds and ulcers. The innovation is based on the extraction, characterization, and application of collagen (CO)/chitosan-glucan complex hollow fibers (CSGC)/aloe vera (AV) as a novel FWS. For the first time, specific hollow fibers were extracted with controlled inner (500-900 nm)/outer (2-3 µm) diameters from mycelium of Schizophyllum commune. Further on, research and evaluation of morphology, hydrolytic stability, and swelling characteristics of CO/CSGC@AV were carried out. The obtained FWS showed high hydrolytic stability with enhanced swelling characteristics compared to native collagen. The hemostatic effect of FWS increased significantly in the presence of CSGC, compared to native CO and displayed excellent biocompatibility which was tested by using normal human dermal fibroblast (NHDF). The FWS showed high antibacterial activity against different types of bacteria (positive/negative grams). From in vivo measurements, the novel FWS increased the percentage of wound closure after one week of treatment. All these results imply that the new CO/CSGC@AV-FWD has the potential for clinical skin regeneration and applying for controlled drug release.

Characterization of physicochemical and biological properties of Schizophyllum commune polysaccharide extracted with different methods.

Water-soluble polysaccharides from Schizophyllum commune (SPGs) were extracted using different methods including hot-water extraction (HWE), ultrasound-assisted extraction (UE), microwave-assisted extraction (ME) and high pressure extraction (HPE). The physicochemical characteristics, antioxidant activity and hypoglycemic activity of four different SPGs were investigated. The results indicated that the polysaccharides yield (8.26%), uronic acid content (3.0%), total polysaccharides content (67.96%) and molecular weight (52.7 kDa) of SPGs extracted by HWE were higher than those obtained by other methods. FT-IR spectra and conformation information of four SPGs were similar, however, the color characteristics, the monosaccharide species and molar ratios, the solubility and micro-structure of each SPGs were significantly different. Though the molecular weight (11.0 kDa) of UE-SPGs was lowest, it showed the highest antioxidant activity and hypoglycemic activity. It could be considered that the UE extraction method might be a good potential extraction technique for the SPGs with high bioactivities.

Studies on structural properties and immune-enhancing activities of glycomannans from Schizophyllum commune.

Polysaccharides were extracted from Schizophyllum commune (a common mushroom) and their structural and immune-enhancing properties were investigated. Crude and fractions (F1 and F2) were composed of sugars (50.3-82.8%), proteins (1.46-20.1%), and sulfates (1.33-7.01%). Monosaccharide compositions of Cr and F1 were mainly composed of glucose (75.5% and 88.2%) with small amounts of mannose, galactose and xylose whereas the F2 was mainly composed of manose (55.2%) with minor amounts of galactose, glucose, and xylose. Their immune-enhancing activities were tested using RAW264.7 cells. Proliferation activity of RAW264.7 cells was over 100% after treatment with these polysaccharides. In addition, RAW264.7 cells produced large amounts of nitric oxide and various cytokines by up-regulating mRNA expression levels and the activation of nuclear factor-kappa (NF-κB) and mitogen-activated protein kinase (MAPK) after treatment with these polysaccharides. In addition, RAW264.7 cells were activated mainly through CR-3 and TLR-4 receptors. The backbone of F2 with excellent immune-enhancing activity was mainly linked by (1→3)-linked-mannopyranosyl and (1→2,3)-linked-mannopyranosyl residues.

Crosstalk between Ras and inositol phosphate signaling revealed by lithium action on inositol monophosphatase in Schizophyllum commune.

Mushroom forming basidiomycete Schizophyllum commune has been used as a tractable model organism to study fungal sexual development. Ras signaling activation via G-protein-coupled receptors (GPCRs) has been postulated to play a significant role in the mating and development of S. commune. In this study, a crosstalk between Ras signaling and inositol phosphate signaling by inositol monophosphatase (IMPase) is revealed. Constitutively active Ras1 leads to the repression of IMPase transcription and lithium action on IMPase activity is compensated by the induction of IMPase at transcriptome level. Astonishingly, in S. commune lithium induces a considerable shift to inositol phosphate metabolism leading to a massive increase in the level of higher phosphorylated inositol species up to the inositol pyrophosphates. The lithium induced metabolic changes are not observable in a constitutively active Ras1 mutant. In addition to that, proteome profile helps us to elucidate an overview of lithium action to the broad aspect of fungal metabolism and cellular signaling. Taken together, these findings imply a crosstalk between Ras and inositol phosphate signaling.

Schizophyllum commune induced genotoxic and cytotoxic effects in Spodoptera litura.

In search for ecofriendly alternatives to chemical insecticides the present study was conducted to assess the insecticidal potential of an endophytic fungus Schizophyllum commune and its mechanism of toxicity by studying genotoxic and cytotoxic effects as well as repair potential using Spodoptera litura (Fabricius) as a model. Different endophytic fungi were isolated and tested for their insecticidal potential against S. litura. Among the tested endophytic fungi maximum mortality against S. litura was exhibited by S. commune isolated from Aloe vera. Extended development, reduced adult emergence was observed in larvae fed on diet supplemented with fungal extract. In addition to it the fungus also has propensity to increase oxidative stress which leads to significantly higher DNA damage. The significantly lower frequency of living haemocytes and increased frequency of apoptotic and necrotic cells was also observed in larvae treated with fungal extract. The extent of recovery of damage caused by fungus was found to be very low indicating long term effect of treatment. Phytochemical analysis revealed the presence of various phenolics, terpenoids and protein in fungal extract. Biosafety analysis indicated the non toxic nature of extract. This is the first report showing the insecticidal potential of S. commune and the genotoxic and cytotoxic effects associated with it.

Molecular weight and helix conformation determine intestinal anti-inflammatory effects of exopolysaccharide from Schizophyllum commune.

Intestinal anti-inflammatory activities of exopolysaccharide from S. commune were assessed using dextran sulfate sodium (DSS)-induced colitis in mice model. The changes of molecular weight (MW), atomic force microscope morphology, X-ray diffraction, particle size distribution, and viscosity were recorded after sonication treatment. The results indicated that the triple helical structure of exopolysaccharide was dissociated into single helical structure and random coiled structure by ultrasonication via breaking of inter- and intramolecular hydrogen bonds. The medium (936kDa) and high MW (1437kDa) exopolysaccharide had the mixture of triple helix and single helix conformation, while the low MW (197kDa) exopolysaccharide exhibit random coiled conformation. The intestinal anti-inflammatory activity study showed that oral administration of medium and high MW (1437kDa) exopolysaccharide significantly recovered DSS-induced colitis in inflamed tissues and reduced inflammation induced infiltration of macrophages. These results showed that medium (936kDa) and high MW (1437kDa) exopolysaccharide had intestinal anti-inflammatory activity. The intestinal anti-inflammatory activity of exopolysaccharide was related to helical structure and molecular weight.

Characterization of cellulolytic enzyme system of Schizophyllum commune mutant and evaluation of its efficiency on biomass hydrolysis.

Schizophyllum commune is a basidiomycete equipped with an efficient cellulolytic enzyme system capable of growth on decaying woods. In this study, production of lignocellulose-degrading enzymes from S. commune mutant G-135 (SC-Cel) on various cellulosic substrates was examined. The highest cellulase activities including CMCase, FPase, and ß-glucosidase were obtained on Avicel-PH101 while a wider range of enzymes attacking non-cellulosic polysaccharides and lignin were found when grown on alkaline-pretreated biomass. Proteomic analysis of SC-Cel also revealed a complex enzyme system comprising seven glycosyl hydrolase families with an accessory carbohydrate esterase, polysaccharide lyase, and auxiliary redox enzymes. SC-Cel obtained on Avicel-PH101 effectively hydrolyzed all agricultural residues with the maximum glucan conversion of 98.0% using corn cobs with an enzyme dosage of 5 FPU/g-biomass. The work showed potential of SC-Cel on hydrolysis of various herbaceous biomass with enhanced efficiency by addition external ß-xylosidase.

Characterization of a Basidiomycota hydrophobin reveals the structural basis for a high-similarity Class I subdivision.

Class I hydrophobins are functional amyloids secreted by fungi. They self-assemble into organized films at interfaces producing structures that include cellular adhesion points and hydrophobic coatings. Here, we present the first structure and solution properties of a unique Class I protein sequence of Basidiomycota origin: the Schizophyllum commune hydrophobin SC16 (hyd1). While the core ß-barrel structure and disulphide bridging characteristic of the hydrophobin family are conserved, its surface properties and secondary structure elements are reminiscent of both Class I and II hydrophobins. Sequence analyses of hydrophobins from 215 fungal species suggest this structure is largely applicable to a high-identity Basidiomycota Class I subdivision (IB). To validate this prediction, structural analysis of a comparatively distinct Class IB sequence from a different fungal order, namely the Phanerochaete carnosa PcaHyd1, indicates secondary structure properties similar to that of SC16. Together, these results form an experimental basis for a high-identity Class I subdivision and contribute to our understanding of functional amyloid formation.

Two new diketopiperazines from the tin mine tailings-derived fungus Schizophyllum commune YIM DT 10058.

Two new diketopiperazines, named diphenylalazine C (1) and epicoccin U (2), together with six known compounds (3-8) were isolated from the EtOAc extract of the fermentation broth of the tin mine tailings-derived fungus Schizophyllum commune YIM DT 10058. Their structures were elucidated by detailed analysis of spectroscopic data and comparison with related known compounds. Compounds 1 and 2 exhibited weak antibacterial and cytotoxic activities.

Lignicolous fungi hydrodistilled extracts may represent a promising source of natural phenolics.

In vitro evaluation of total phenolic contents and antiradical activities of the lignicolous fungi Fomes fomentarius and Schizophyllum commune hydrodistilled extracts was the subject of this study. This preliminary screening included four free radical species evaluated by UV-vis (DPPH•, ABTS• and •NO) and EPR (Asc•), respectively. According to the experimental data obtained, both F. fomentarius and S. commune hydrodistilled extracts may be considered as promising sources of phenolic natural products (157 and 138 mg GAE/g d.e., respectively) and other bioactives showing good anti-DPPH (1.31 µg/mL) and anti-Asc (70.40%) radical activities, respectively, at in vitro conditions.

Microbial Glucuronoyl Esterases: 10 Years after Discovery.

A carbohydrate esterase called glucuronoyl esterase (GE) was discovered 10 years ago in a cellulolytic system of the wood-rotting fungus Schizophyllum commune Genes coding for GEs were subsequently found in a number of microbial genomes, and a new family of carbohydrate esterases (CE15) has been established. The multidomain structures of GEs, together with their catalytic properties on artificial substrates and positive effect on enzymatic saccharification of plant biomass, led to the view that the esterases evolved for hydrolysis of the ester linkages between 4-O-methyl-d-glucuronic acid of plant glucuronoxylans and lignin alcohols, one of the crosslinks in the plant cell walls. This idea of the function of GEs is further supported by the effects of cloning of fungal GEs in plants and by very recently reported evidence for changes in the size of isolated lignin-carbohydrate complexes due to uronic acid de-esterification. These facts make GEs interesting candidates for biotechnological applications in plant biomass processing and genetic modification of plants. This article is a brief summary of current knowledge of these relatively recent and unexplored esterases.