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.

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.

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).

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.

Phenolic acids isolated from the fungus Schizophyllum commune exert analgesic activity by inhibiting voltage-gated sodium channels.

The present study was designed to search for compounds with analgesic activity from the Schizophyllum commune (SC), which is widely consumed as edible and medicinal mushroom world. Thin layer chromatography (TLC), tosilica gel column chromatography, sephadex LH 20, and reverse-phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify compounds from SC. Structural analysis of the isolated compounds was based on nuclear magnetic resonance (NMR). The effects of these compounds on voltage-gated sodium (NaV) channels were evaluated using patch clamp. The analgesic activity of these compounds was tested in two types of mouse pain models induced by noxious chemicals. Five phenolic acids identified from SC extracts in the present study included vanillic acid, m-hydroxybenzoic acid, o-hydroxybenzeneacetic acid, 3-hydroxy-5-methybenzoic acid, and p-hydroxybenzoic acid. They inhibited the activity of both tetrodotoxin-resistant (TTX-r) and tetrodotoxin-sensitive (TTX-s) NaV channels. All the compounds showed low selectivity on NaV channel subtypes. After intraperitoneal injection, three compounds of these compounds exerted analgesic activity in mice. In conclusion, phenolic acids identified in SC demonstrated analgesic activity, facilitating the mechanistic studies of SC in the treatment of neurasthenia.

Selective isolation of hydrophobin SC3 by solid-phase extraction with polytetrafluoroethylene microparticles and subsequent mass spectrometric analysis.

Hydrophobins are small proteins that play a role in a number of processes during the filamentous fungi growth and development. These proteins are characterized by the self-assembly of their molecules into an amphipathic membrane at hydrophilic-hydrophobic interfaces. Isolation and purification of hydrophobins generally present a challenge in their analysis. Hydrophobin SC3 from Schizophyllum commune was selected as a representative of class I hydrophobins in this work. A novel procedure for selective and effective isolation of hydrophobin SC3 based on solid-phase extraction with polytetrafluoroethylene microparticles loaded in a small self-made microcolumn is reported. The tailored binding of hydrophobins to polytetrafluoroethylene followed by harsh elution conditions resulted in a highly specific isolation of hydrophobin SC3 from the model mixture of ten proteins. The presented isolation protocol can have a positive impact on the analysis and utilization of these proteins including all class I hydrophobins. Hydrophobin SC3 was further subjected to reduction of its highly stable disulfide bonds and to chymotryptic digestion followed by mass spectrometric analysis. The isolation and digestion protocols presented in this work make the analysis of these highly hydrophobic and compact proteins possible.

Endothelium-dependent and -independent relaxation of rat aorta induced by extract of Schizophyllum commune.

Schizophyllum commune (SC) is widely consumed by Chinese, especially in southern part of China. The aim of the present study was to assess the extract of SC on vascular tone and the mechanisms involved. Experiments were performed on aorta of 18-week-old male Sprague-Dawley rats. Dried SC was extracted with 50% ethanol, 90% ethanol and deionized water, respectively. The effects of SC on the isometric tension of rat aortic rings were measured. Protein expression for the endothelial nitric oxide synthase (eNOS) was also determined in the primarily cultured rat aortic arterial endothelial cells (RAECs). The results showed that the water extract of SC induced a marked relaxation in aortic rings with or without endothelium. After the pretreatments of N(ω)-nitro-l-arginine methyl ester, indomethacin, RP-cAMP, and methylene blue, the SC-induced relaxation was significantly decreased. In addition, the contraction due to Ca(2+) influx and intracellular Ca(2+) release was also inhibited by SC. Furthermore, expression of the eNOS protein was significantly elevated in RAECs after treatment of SC. In conclusion, the water extract of SC induces an endothelium-dependent and -independent relaxation in rat aorta. The relaxing effect of SC involves the modulation of NO-cGMP-dependent pathways, PGI2-cAMP-depedent pathways, Ca(2+) influx though calcium channels and intracellular Ca(2+) release.

The antitumor activity of hydrophobin SC3, a fungal protein.

The use of mushroom extracts has been common practice in traditional medicine for centuries, including the treatment of cancer. Proteins called hydrophobins are very abundant in mushrooms. Here, it was examined whether they have antitumor activity. Hydrophobin SC3 of Schizophyllum commune was injected daily intraperitoneally starting 1 day after tumor induction in two tumor mouse models (sarcoma and melanoma). SC3 reduced the size and weight of the melanoma significantly, but the sarcoma seemed not affected. However, microscopic analysis of the tumors 12 days after induction revealed a strong antitumor effect of SC3 on both tumors. The mitotic activity of the tumor decreased 1.6- (melanoma) to 2.3-fold (sarcoma), while the vital mass decreased 2.3- (melanoma) to 4.3-fold (sarcoma) compared to the control. Treatment did not cause any signs of toxicity. Behavior, animal growth, and weight of organs were similar to animals injected with vehicle, and no histological abnormalities were found in the organs. In vitro cell culture studies revealed no direct cytotoxic effect of SC3 towards sarcoma cells, while cytotoxic activity was observed towards melanoma cells at a high SC3 concentration. Daily treatment with SC3 did not result in detectable levels of anti-SC3 antibodies in the plasma. Instead, a cellular immune response was observed. Incubation of spleen cells with SC3 resulted in a 1.5- to 2.5-fold increase in interleukin-10 and TNF-α mRNA levels. In conclusion, the nontoxic fungal hydrophobin SC3 showed tumor-suppressive activity possibly via immunomodulation and may be of benefit as adjuvant in combination with chemotherapy and radiation.

Oxidative stability of sunflower oil supplemented with medicinal split gill mushroom, Schizophyllum commune Fr.:Fr. extract during accelerated storage.

The oxidative stability of sunflower oil supplemented with medicinal split gill mushroom, Schizophyllum commune's crude extract (CE), the formic acid (FA) fraction and semipurified subfractions (SF) II and IV were tested, compared to BHA and alpha-tocopherol, by measuring their peroxide value, iodine value, p-anisidine value, thiobarbituric acid-reactive substances, and free fatty acid content. Their total phenolic content (TPC), 2,2-diphenyl-1-picryhydrazyl (DPPH) radical scavenging, and ferric reducing/antioxidant power (FRAP) were also evaluated. FA and CE exhibited highest DPPH* scavenging, while FA and SFIV showed the highest FRAP; TPC was found to be highest in CE, FA, and SFIV. BHA and alpha-tocopherol are more protective in stabilizing the sunflower oil; SFII and SFIV had short-term protective effect in secondary oxidation for 1 year, while CE and FA retarded secondary oxidation and extended the shelf life 1 1/2 years and 2 years, respectively. HPLC-DAD analysis found (+)-catechin in Sch. commune's extracts. Sch. commune's extracts did not show similar retardation of lipid oxidation in sunflower oil as compared to alpha-tocopherol and BHA at the 200 ppm level. However, the higher concentration of Sch. commune's extract that provided the protective effect in stabilizing sunflower oil can be further studied.

Protease purified from Schizophyllum commune culture broth digests fibrins without activating plasminogen.

Schizophyllum commune is a widely distributed mushroom used as an herbal medicine and an ingredient in healthy food. In this study, a protease from a fermented culture broth of S. commune demonstrated strong fibrinolytic and fibrinogenolytic activities. This fibrinolytic protease showed a suppression effect in blood coagulation in co-incubation with rat citrated blood through thromboelastographic analysis. The protease suppressed aggregation of fibrin (ogen), but not the platelets, in clotting formation and significantly decreased the clot strength. We also found very little potency in this protease to activate plasminogen, thus it exhibits the potential for an ideal fibrinolytic candidate for therapeutic applications in the future.

Cerebrosides A and C, sphingolipid elicitors of hypersensitive cell death and phytoalexin accumulation in rice plants.

When plants interact with certain pathogens, they protect themselves by generating various chemical and physical barriers called the hypersensitive response. These barriers are induced by molecules called elicitors that are produced by pathogens. In the present study, the most active elicitors of the hypersensitive response in rice were isolated from the rice pathogenic fungus Magnaporthe grisea, and their structures were identified as cerebrosides A and C, sphingolipids that were previously isolated as inducers of cell differentiation in the fungus Schizophyllum commune. Treatment of rice leaves with cerebroside A induced the accumulation of antimicrobial compounds (phytoalexins), cell death, and increased resistance to subsequent infection by compatible pathogens. The degradation products of cerebroside A (fatty acid methyl ester, sphingoid base, and glucosyl sphingoid base) showed no elicitor activity. Hydrogenation of the 8E-double bond in the sphingoid base moiety or the 3E-double bond in the fatty acid moiety of cerebroside A did not alter the elicitor activity, whereas hydrogenation of the 4E-double bond in the sphingoid base moiety led to a 12-fold decrease in elicitor activity. Furthermore, glucocerebrosides from Gaucher's spleen consisting of (E)-4-sphingenine and cerebrosides from rice bran mainly consisting of (4E,8E)-4,8-sphingadienine and (4E,8Z)-4,8-sphingadienine showed no elicitor activity. These results indicate that the methyl group at C-9 and the 4E-double bond in the sphingoid base moiety of cerebrosides A and C are the key elements determining the elicitor activity of these compounds. This study is the first to show that sphingolipids have elicitor activity in plants.