Chemical Components Analyzed by UPLC-QTOF-MS and In Vitro Antitumor Effects of the Chestnut Mushroom Pholiota adiposa (Agaricomycetes).

We investigated the chemical constituents and anti-tumor activity of cultivated Pholiota adiposa in vitro using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. HepG-2, A549, HeLa, and MCF-7 cells, which are 4 kinds of human cancer cell lines, were cultured in vitro, treated with different concentrations of the ethanol extract of Ph. adiposa (EPA), and cytotoxicity was determined using the cell counting kit-8 assay. Flow cytometry was used to analyze the apoptosis of HepG-2 cells via annexin V-fluorescein isothiocyanate/propidium iodide double staining. Expression levels of apoptosis-associated proteins were determined via Western blotting analysis. Thirty-five components were consistent with those recorded in the chemical composition database, with sterols, fatty acids, and polysaccharide compounds accounting for a relatively high proportion. EPA showed the strongest cytotoxicity against HepG-2 cells, increasing the apoptosis rate up to 23.71 ± 1.59% at a concentration of 50 µg/mL. Ph. adiposa has various functional chemical constituents and potential anti-tumor applications. We found that the functional constituents exerted anti-tumor activity by inducing apoptosis. Furthermore, the expression levels of BCL-2-associated X were increased, whereas those of BCL-2 were decreased in cells after treatment with EPA. These results suggest that EPA induces HepG-2 cell apoptosis via a caspase-mediated pathway.

Anti-diabetic effects of fungal Ergosta-4, 6, 8(14), 22-tetraen-3-one from Pholiota adiposa.

OBJECTIVE: Pholiota adiposa is a valuable edible and medicinal fungus. In this research, Ergosta-4, 6, 8(14), 22-tetraen-3-one (ETO) was obtained from Pholiota adiposa which is the first time to study the anti-diabetic and related mechanism. METHODS: Ergosta-4, 6, 8 (14), 22-tetraen-3-one (ETO) was defined by IR and NMR. Relevant biochemical indicators were detected by ELISA assay, hematoxylin-eosin staining (H & E), fasting glucose levels (FBG), oral glucose tolerance test (OGTT), tissue homogenate biochemical measurements, immunohistochemical staining and western blot. RESULTS: In this research, the ETO treatment groups exhibited a significant reduction in fasting blood glucose (FBG) levels, the High dose group (HD) was about 10 mmol/L lower than the diabetic control group (DC), and increase in body weight, the HD group weighed about 5 g more than the DC group on average. Also, the levels of triacylglycerol (TG), total cholesterol (TC), and low-density lipoprotein-cholesterol (LDL-C) were found to be decreased, while the levels of high-density superoxide dismutase (SOD), lipoproteincholesterol (HDL-C), catalase (CAT), and glutathione peroxidase (GSH-Px) were increased in the ETO treatment groups. The pancreatic and liver sections of diabetic control group (DC) exhibited several histopathological changes, but the ETO treatment groups exhibited improvements. ETO treatment led to the significant restoration of islet morphology and function. Moreover, the results of the western blot analysis indicate that ETO could be used for the treatment of diabetes, since it modifies part of the IRS1 / PI3K / AKT signaling pathway.

New polyketides from the basidiomycetous fungus Pholiota sp.

Two new polyketides, pholiotones B and C (1 and 2), and four known compounds, trichodermatide D (3), vermistatin (4), dehydroaltenuene A (5) and terpestacin (6) were isolated from the crude extract of Pholiota sp. Their structures were identified by NMR and MS spectroscopic data. The absolute configurations of compounds 1 and 2 were elucidated by modified Mosher's method, electronic circular dichroism (ECD) calculations and 13C NMR calculations as well as DP4+ probability analyses. All the compounds were evaluated for their antifungal and cytotoxicity.

Core fucose-specific Pholiota squarrosa lectin (PhoSL) as a potent broad-spectrum inhibitor of SARS-CoV-2 infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein (S protein) is highly N-glycosylated, and a "glycan shield" is formed to limit the access of other molecules; however, a small open area coincides with the interface to the host's receptor and also neutralising antibodies. Most of the variants of concern have mutations in this area, which could reduce the efficacy of existing antibodies. In contrast, N-glycosylation sites are relatively invariant, and some are essential for infection. Here, we observed that the S proteins of the ancestral (Wuhan) and Omicron strains bind with Pholiota squarrosa lectin (PhoSL), a 40-amino-acid chemically synthesised peptide specific to core-fucosylated N-glycans. The affinities were at a low nanomolar level, which were ~ 1000-fold stronger than those between PhoSL and the core-fucosylated N-glycans at the micromolar level. We demonstrated that PhoSL inhibited infection by both strains at similar submicromolar levels, suggesting its broad-spectrum effect on SARS-CoV-2 variants. Cryogenic electron microscopy revealed that PhoSL caused an aggregation of the S protein, which was likely due to the multivalence of both the trimeric PhoSL and S protein. This characteristic is likely relevant to the inhibitory mechanism. Structural modelling of the PhoSL-S protein complex indicated that PhoSL was in contact with the amino acids of the S protein, which explains the enhanced affinity with S protein and also indicates the significant potential for developing specific binders by the engineering of PhoSL.

Rheological properties of polysaccharides from Pholiota nameko with different temperature extraction: Concentration, pH, temperature, and saltion.

Cold and hot water extracted polysaccharides (CW-PNPs and HW-PNPs) were isolated from Pholiota nameko. The rheological properties of PNPs were investigated by steady shear and oscillatory rheological measurements. The PNPs exhibited typical non-Newtonian and shear-thinning behavior, which are affected by PNP concentration, temperature, pH value, salt ion, and concentration. Specifically, the apparent viscosity of the two PNPs solutions at concentration of 1% (w/w) was shown as HW-PNPs > CW-PNPs. The apparent viscosity of PNPs decreases under acidic and alkaline conditions and when the temperature rises; K+ and Na+ cause the apparent viscosity of CW-PNPs to decrease, while Ca2+ and Al3+ are opposite. The addition of four different salt ions all caused the apparent viscosity of the HW-PNPs to decrease. The results of dynamic rheological experiments show that G' and G″ showed slightly frequency dependency with G' exceeding G″ throughout the accessible range of frequency for CW-PNPs and HW-PNPs.

Structural analysis and in vitro antitumor effect of polysaccharides from Pholiota adiposa.

Pholiota adiposa is an edible chestnut mushroom with many health benefits, such as antioxidant and anticancer activity. In this paper, polysaccharides were extracted from Pholidota adiposa using an acid extraction process. The crude polysaccharide was purified using DEAE-cellulose chromatography, and two polysaccharide fractions of SPAP2-1 and SPAP2-2 were obtained. The structure was characterized using UV, GPC, GC, FT-IR, methylation, and NMR analysis. Monosaccharide component analysis indicated that SPAP2-1 (19 kDa) and SPAP2-2 (20 kDa) contained mannose, glucose, and galactose with different molecular ratios. Their antitumor effects were investigated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium (MTT) assay, Annexin V-fluorescein isothiocyanate (FITC), propidium iodide (PI) staining, and flow cytometry. By analyzing the changes in the cells, SPAP2-1 caused damage and changed the proliferation rate of HeLa cells. SPAP2-1 showed strong interference to the cell cycle of HeLa cells and induced cell apoptosis. Overall, these results suggested that polysaccharides from Pholiota adiposa, especially SPAP2-1, may have the potential to be used as a tumor cell inhibitor, which needs further study.

Influence of Pholiota adiposa on gut microbiota and promote tumor cell apoptosis properties in H22 tumor-bearing mice.

Hepatocellular carcinoma (HCC) is a common type of cancer-prevalent worldwide-and one of the causes of cancer-related deaths. In this study, ethanol extracts from Pholiota adiposa (EPA) were used to identify possible targets for HCC treatment and their effects on intestinal microflora were analyzed. Methods: Male mice were randomly assigned to groups-the model group, cyclophosphamide (25 mg/kg/d), and EPA groups, in which the mice were categorized based on the different concentrations of each compound (100, 200, and 300 mg/kg/day). Relevant biochemical indicators were detected using ELISA, H&E staining, and TUNEL assay. Four tumor apoptosis-related proteins and genes, Cleaved Caspases, BAX, Bcl-2, and VEGF, were detected by immunohistochemical staining, western blotting, and RT-PCR. The total genomic DNA was obtained from the contents of the small intestine and colon and was sequenced. The V3 + V4 regions of bacterial 16 s rDNA (from 341 to 806) were amplified. Results: The tests revealed that EPA exhibited antitumor activity in vivo by promoting apoptosis and inhibiting angiogenesis. Moreover, EPA treatment could increase beneficial and decrease harmful microflorae. These results demonstrate that EPA may be a potential therapy for HCC.

Study on the physicochemical properties and immunomodulatory anti-tumor effect of the Pholiota adiposa polysaccharide.

In this study, the extraction, purification, physical and chemical properties, and biological activity of the Pholiota adiposa (PAP) polysaccharide were investigated. One fraction (PAP-1a) of Pholiota adiposa polysaccharides was isolated using DEAE Sepharose™ Fast Flow and Sephacryl™ S-300 High-Resolution columns. The HPLGPC results revealed that the molecular weight of PAP-1a was 16.453 kDa. PAP-1a was composed of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, and fucose and their molar % was 33.41, 0.53, 1.33, 0.07, 0.27, 5.28, 38.31, 0.83, 18.04 and 2.23, respectively. PAP-1a could activate macrophages to secrete NO and cytokines such as TNF-a, IL-6, and IL-12p70. When hepatocellular carcinoma cells (HCCs) and macrophages were co-cultured, it was observed that PAP-1a inhibited the growth of Hep-G2, Hep-3B, and Huh7 via immunoregulation. It triggered cell apoptosis by blocking the cell cycle in the G0/G1 stage. Furthermore, PAP-1a had no direct cytotoxicity against the hepatocyte cell line L02 and macrophages RAW264.7.

Green synthesis and characterization of gold nanoparticles from Pholiota adiposa and their anticancer effects on hepatic carcinoma.

Gold nanoparticles (AuNPs) were successfully fabricated by Pholiota adiposa polysaccharide (PAP-1a) without employing any other chemicals. The physical and chemical properties of PAP-AuNPs were determined using transmission electron microscopy (TEM), dynamic light scattering (DLS), energy-dispersive X-ray spectroscopy (EDXR), Fourier-transform infrared spectroscopy (FT-IR), and atomic force microscopy (AFM). In an attempt to analyze the immune regulation, antitumor effect, and biological safety, the production of NO and TNF-α, IL-12p70, and IL-1ß from RAW264.7 as well as the proliferation of RAW264.7 were detected in vitro. Flow cytometry was conducted to determine the ratio of the CD4+/CD8+ cell in peripheral blood and immunohistochemical analysis involving hematoxylin and eosin (H&E) and proliferating cell nuclear antigen (PCNA) staining were conducted in vivo. The results of this study showed that PAP-AuNPs had a significantly improved immune regulation and anti-tumor effect in comparison to PAP-1a alone. PAP-AuNPs showed no toxicity both in vivo and in vitro. This study demonstrates a useful application of PAP-AuNPs as a novel nanomedicine for hepatic carcinoma.

Structure characterization and antioxidant activity of carboxymethylated polysaccharide from Pholiota nameko.

The objective of this paper was to prepare the carboxymethyl polysaccharide of Pholiota nameko (CPPN) and further evaluate its structural characteristics, water solubility, and antioxidant activities. The optimal carboxymethylation conditions were as follows: 20% NaOH concentration, 1.5 hr alkalization, 4 hr etherification time, and 60°C with 2 g chloroacetic acid. Under these conditions, a substitution degree of 0.72 and a yield of 88.3% were achieved. The results of NMR and FT-IR spectra showed that carboxymethylation modification was successful. The protein, total sugar, and total phenol content decreased, while the molecular weight increased, and the monosaccharide composition content and surface structure changed upon carboxymethylation. Compared with unmodified polysaccharides, CPPN had significant antioxidant activity and water solubility. Thus, this study could provide a potential antioxidant for functional foods and pharmaceuticals. PRACTICAL APPLICATIONS: Pholiota nameko is an edible and medicinal mushroom, rich in carbohydrates, cellulose, and minerals. Pharmacological studies have shown that P. nameko polysaccharides have many medicinal effects on the human body. In this study, CPPN was synthesized by carboxymethylation of P. nameko polysaccharides. Through structural characterization, antioxidant activity, and solubility studies, carboxymethylated polysaccharides shown significant antioxidant properties and increased solubility. Therefore, this research lays the foundation for the high-value application of P. nameko polysaccharides in functional food and pharmaceutical industry.

Chemical properties and anti-fatigue effect of polysaccharide from Pholiota nameko.

The aim of this study was to explore the primary chemical properties and anti-fatigue effect in vivo of Pholiota nameko polysaccharide (PNP). Through UV-visible spectrum, the absorption peaks of proteins, nucleic acids and pigments were not found. The organic functional groups of polysaccharides (3,289.97, 1,584.72, and 1,045.23 cm-1 so on) were measured by IR spectroscopy. The PNP was a semi-crystalline or non-crystalline substance, possessed a three-dimensional lump structure with a smooth, dense surface and amorphous structure according to the scanning electron microscopy and XRD images. Moreover, the PNP was chain or bright-spot structures formed by the entanglement of multiple polysaccharide fibers on the basis of atomic force microscopy. The results of anti-fatigue suggested the PNP could significantly extend the forced swim time from 121.58 ± 18.48 and 101.91 ± 14.27 min to 154.95 ± 24.26 and 134.13 ± 25.71 min in male and female mice respectively. The LDH activity was up to 31.68 ± 4.60 U/ml in male mice and 29.49 ± 5.12 U/ml in female mice. Meanwhile, the Ca2+ -Mg2+ -ATPase activity was reached to 2.49 ± 0.41 µmol/(mg·h) in male mice and 2.44 ± 0.29 µmol/(mg·h) in female mice. The SOD activity was increased to 5.92 ± 1.19 U/ml in male mice and 5.89 ± 0.98 U/ml in female mice, while the MDA content was decreased to 2.24 ± 0.34 nmol/mg in male mice and 2.02 ± 0.41 nmol/mg in female mice. These results showed a theoretical basis for application of the PNP in food and pharmacy as a natural physical strengthening substance. PRACTICAL APPLICATIONS: Fatigue affects physical and mental health in vivo, which resulted in negative effects on everyday tasks, leisure activities, cognitive and behavioral performances and is very common in modern life. Therefore, this study was designed to explore the primary chemical properties and research the anti-fatigue effects of Pholiota nameko polysaccharide (PNP) in mice. And then, it would be a reference for the development and utilization of PNP as a kind of healthy food on sub-health.

Serum core-type fucosylated prostate-specific antigen index for the detection of high-risk prostate cancer.

It is known that core-type fucosylation is higher in prostate cancer cells than in other cancer cell types and is associated with high-risk prostate cancer. Here, we developed an automated microcapillary electrophoresis-based immunoassay system for measuring serum core-type fucosylated prostate-specific antigen (PSA) and evaluated whether the serum fucosylated PSA index (FPI) can detect high-risk prostate cancer. Core-type fucosylated-free PSA was measured by our automated microcapillary electrophoresis-based immunoassay system with Pholiota squarrosa lectin. The FPI was calculated from total PSA and the percentage of fucosylated-free PSA. The optimum model to predict Gleason grade (GG) ≥2 was constructed by multivariate logistic regression analysis. Discrimination was assessed by determining the area under the receiver operator characteristic curve (AUC). The study included 252 men who underwent prostate needle biopsy due to elevated serum PSA levels (4-20 ng/mL), including 138 with GG ≥2. A higher FPI was significantly associated with GG (P < .0001). Multivariate logistic regression analysis showed that age, prostate volume and FPI were significant predictors of GG ≥2. The AUC of FPI and the model were 0.729 (95% confidence interval [CI]: 0.668-0.790) and 0.837 (95% CI: 0.788-0.886), respectively, compared to 0.629 (95% CI: 0.561-0.698) for PSA. Decision curve analysis showed the superior benefit of FPI and the model when compared to PSA. In a cohort with serum PSA levels <20 ng/mL, FPI could differentiate high-risk prostate cancer from biopsy-negative or low-risk prostate cancer. Therefore, FPI could be a useful adjunct in prostate biopsy counseling for men with abnormal PSA levels.

Glycan array analysis of Pholiota squarrosa lectin and other fucose-oriented lectins.

The α(1,6)fucose residue attached to the N-glycoprotein core is suspected to play an essential role in the progression of several types of cancer. Lectins remain the first choice for probing glycan modifications, although they may lack specificity. Thus, efforts have been made to identify new lectins with a narrower core fucose (CF) detection profile. Here, we present a comparison of the classical Aleuria aurantia lectin (AAL), Lens culinaris agglutinin (LCA) and Aspergillus oryzae lectin (AOL) with the newer Pholiota squarrosa lectin (PhoSL), which has been described as being specific for core fucosylated N-glycans. To this end, we studied the binding profiles of the four lectins using mammalian glycan arrays from the Consortium of Functional Glycomics. To validate their glycan specificity, we probed AOL, LCA and PhoSL in western-blot assays using protein extracts from eight common colorectal cancer (CRC) lines and colorectal biopsies from a small cohort of patients with CRC. The results showed that (i) LCA and PhoSL were the most specific lectins for detecting the presence of CF in a concentration-dependent manner; (ii) PhoSL exhibited the highest N-glycan sequence restriction, with preferential binding to core fucosylated paucimannosidic-type N-glycans, (iii) the recognition ability of PhoSL was highly influenced by the presence of terminal N-acetyl-lactosamine; (iv) LCA bound to paucimannosidic, bi-antennary and tri-antennary core fucosylated N-glycans and (v) AOL and AAL exhibited broader specificity towards fucosylation. Together, our results support the choice of LCA as the most appropriate lectin for CF detection, as validated in protein extracts from CRC cell lines and tissue specimens from patients with CRC.

A phylogenetic assessment of Pholiota and the new genus Pyrrhulomyces.

Multigene data sets were assembled to evaluate the phylogeny of species attributed to the genus Pholiota sensu A.H. Sm. & Hesler. This effort included generation of just more than 200 new sequences from 19 type collections of Pholiota and recent samples from East Asia. Phylogenetic analyses reinforced the autonomous phylogenetic positions of pholiotoid taxa in the genera Flammula (Hymenogastraceae) and Kuehneromyces (Strophariaceae). Samples of Pholiota astragalina from diverse geographic regions split into two species-level lineages but occupied an isolated phylogenetic position apart from Pholiota sensu stricto. The new genus Pyrrhulomyces is described to accommodate P. astragalina and a new cryptic species from the Southern Appalachians, Pyrrhulomyces amariceps. Pyrrhulomyces is distinguished from other genera of Strophariaceae by the blackening basidiomata with a bitter taste, smooth basidiospores without a germ pore under light microscopy, presence of pleurochrysocystidia, an ixocutis, rugulose spore ornamentation under scanning electron microscope (SEM), and association with late stages of conifer wood decay. Pholiota subochracea was found to be sister to a clade containing samples of Hypholoma and Bogbodia, but this portion of the Strophariaceae will require further taxon and gene sampling to resolve relationships between these three taxa. Pholiota sensu stricto comprised at least two major groups, but several residual poorly placed lineages were also noted depending on the data set analyzed. New combinations are made in the genera Flammula, Kuehneromyces, and Stropharia for three species of Pholiota-P. abieticola, P. obscura, and P. scabella, respectively, based on molecular annotation of type collections. Overall, 20 new synonymies are proposed, mostly in Pholiota. Illustrations of Pyrrhulomyces are provided along with a key to genera of Strophariaceae and Hymenogastraceae.

Increased antioxidant activity and improved structural characterization of sulfuric acid-treated stepwise degraded polysaccharides from Pholiota nameko PN-01.

The aim of this study was to investigate sulfuric acid degradation of the Pholiota nameko polysaccharide (AIPS-1). Three stepwise degraded polysaccharides (AIPS-2, AIPS-3, and AIPS-4) were obtained by sequentially increasing the strength of sulfuric acid treatment. Structural characterization showed that sulfuric acid treatment significantly decreased molecular weight, increased the content of uronic acid and changed the molar ratio of monosaccharide composition, while the major functional groups and the triple helical conformation of polysaccharides did not change significantly. In vitro experiments proved that the antioxidation ability of the stepwise degraded polysaccharides gradually increased (AIPS-1 < AIPS-2 < AIPS-3 < AIPS-4). An oxidative stress zebrafish model was established, which demonstrated that the ability of AIPS-3 and AIPS-4 to scavenge free radicals in zebrafish was significantly improved compared to AIPS-1. In conclusion, sulfuric acid treatment is an effective method for improving the antioxidant activity of polysaccharides, and increased antioxidant activity was closely related to the changes in their structural characteristics.

An ultrasmall chitosan nanosphere encapsulating carbon dots and rhodamine B as a ratiometric probe for the determination of Hg2.

Hg2+-sensitive carbon dots (CDs) were synthesized by microwave-assisted pyrolysis of citric acid, sodium fluoride, and urea. The CDs as a signal report unit and rhodamine B (RhB) as a reference were then encapsulated in a nanosphere of chitosan assembled by a nonsolvent-induced chitosan colloidal formation and in situ cross-linking to construct a ratiometric probe for Hg2+ (chitosan-CDs-RhB). Interestingly, without any assistance from acids to improve the solubility of chitosan, the nanosphere containing CDs and RhB had an ultrasmall size of 9.7 nm with only approximately 1.1-nm-thick layers of chitosan enclosing one dot. In order to keep the residual functional groups on the nanosphere from compromising the fluorescence response of CDs to Hg2+, Co2+ was used as a fluorescently intact metal ion to saturate the functional groups. The saturated chitosan-CDs-RhB was thus potentially developed for determining Hg2+ in the fruit bodies and mycelia of edible and medicinal fungi. Limits of detection (LODs) of 2.24, 5.29, and 2.03 µM and recoveries in the ranges 98.3 to 101.8%, 99.5 to 104.6%, and 97.4 to 100.9% were estimated for the determination of Hg2+ in the fruit bodies of Pleurotus ostreatus, Lentinus edodes, and Hypsizygus marmoreus, respectively. Chitosan-CDs-RhB was further developed as a fluorescent ratiometric probe for quantitatively determining intracellular Hg2+ in fungal mycelia with a linear calibration curve of RIgreen/Ired = - 0.145c + 1.69 within the range 0.013 to 0.356 µg g-1. Graphical abstract An ultrasmall chitosan nanosphere encapsulating carbon dots and rhodamine B as a ratiometric probe for the determination of Hg2+.

A novel antitumor protein from the mushroom Pholiota nameko induces apoptosis of human breast adenocarcinoma MCF-7 cells in vivo and modulates cytokine secretion in mice bearing MCF-7 xenografts.

Breast cancer is the most common malignant tumor in women and one of the three most common cancers worldwide. It is a life-threatening disease among women and the leading cause of death among women. New drugs or new drug translations and laboratory clinical studies are ongoing. A new antitumor protein (PNAP) purified from edible fungus Pholiota nameko has potential for treating breast cancer. We have previously found that PNAP exhibits anti-proliferative and apoptosis-inducing activities in a human breast cancer cell line (MCF-7). In this study, we constructed a BALB/c mouse model of MCF-7 tumor xenografts. In vivo experiments show that PNAP can effectively inhibit the malignant proliferation of MCF-7 solid tumors. This is because PNAP can successfully activate the death receptor pathway and mitochondrial apoptosis pathway of MCF-7 tumor cells in vivo, and induce tumor cells to wither. It is estimated that PNAP may also have an immunoregulatory ability to indirectly inhibit malignant proliferation of tumors. We also found that PNAP may also have the immunomodulatory ability to indirectly inhibit the malignant proliferation of tumors, which can shift the balance of Th1/Th2 to Th1 and eventually inhibit the growth of tumors. The study reveals a new therapeutic approach for breast cancer patient.

Pholiotone A, a new polyketide derivative from Pholiota sp.

Pholiotone A (1), a new polyketide derivative, with tetrahydrobenzofuran-4(2H)-one skeleton, together with four known compounds, trichodermatides A (2) and B (3) and koninginins B (4) and E (5), were isolated from the crude extract of Pholiota sp. The structures of all the isolated compounds were determined mainly by NMR experiments, the modified Mosher method and electronic circular dichroism (ECD) calculations. The antifungal and cytotoxicity of all isolates were evaluated.

One new sterpurane sesquiterpene from cultures of the basidiomycete Pholiota nameko.

One new sterpurane sesquiterpene (1), named (3R,6S,7S,8R,10S)-3,7,14-trihydroxy-1-sterpurene was isolated from cultures of the basidiomycete Pholiota nameko. The structure of new compound was elucidated by extensive spectroscopic. Additionally, a single crystal X-ray diffraction not only confirmed the structure, but also determined the absolute configuration of the new compound. The compound was evaluated for cytotoxicity against five human cancer cell lines, but no significant cytotoxicity were found (IC50 values > 40 µM).

Effect of ultrasound-assisted extraction on physicochemical properties and TLR2-affinity binding of the polysaccharides from Pholiota nameko.

The effect of the ultrasonic treatment on the physicochemical properties, oil-holding capacities, foaming capacities, and TLR2-affinity of the polysaccharides from Pholiota nameko (PNPS), was evaluated. Compared with the protein content of PNPS before ultrasonic treatment, the protein content of PNPS all presented a decrease under different ultrasonic conditions. The viscosity of PNPS showed a decrease when the ultrasonic intensity was strong enough, as well as the molecular weight. The oil-holding capacity and the foaming capacity of PNPS showed a continuous increasing trend with the increase of the ultrasonic treatment time under a set ultrasonic power of 400 W. Further, the ultrasonic operation could induce the decrease of the affinity binding between PNPS and the receptor proteins TLR2. These data confirmed that applying the ultrasonic treatment could obtain PNPS with high carbohydrate contents, low viscosity and low TLR2-affinity under proper ultrasonic condition.