Effect of Flammulina velutipes polysaccharides on endoplasmic reticulum stress-mediated apoptosis by activating PLC-IP3 pathway in HepG2 cells.

Flammulina velutipes polysaccharides (FVP) have been proven to induce apoptosis in HepG2 cells. It is well known that endoplasmic reticulum stress (ERS) is involved in apoptosis. However, ERS mediates FVP-induced apoptosis in HepG2 cells remains unclear. In our study, the results indicated that FVP caused ERS in HepG2 cells. They showed that FVP were water-soluble polysaccharides with the weight average molecular weight of 1972 kDa, which were mainly composed of mannose, gluconic acid, glucose, galactose, xylose and fructose in a molar ratio of 6.6 : 1.3 : 79.9 : 7.4 : 3.4 : 1.5. After FVP treatment, the expression levels of genes and proteins related to ERS were upregulated. The inhibition of ERS by 4-phenylbutyric acid (4-PBA) pretreatment could significantly reduce the role of FVP in inducing apoptosis. We further found the results of immunofluorescence and flow cytometry showing that Ca2+ in the ERS leaked out, and the intracellular Ca2+ concentration increased after FVP treatment. The pretreatment with the phospholipase C (PLC) inhibitor U73122 proved that FVP caused excessive intracellular Ca2+ concentration by activating the phospholipase C-inositol-1,4,5-triphosphate (PLC-IP3) pathway, resulting in ERS, and ultimately leading to apoptosis. In summary, our results indicated that FVP induced ERS-mediated apoptosis by activating PLC-IP3 pathway in HepG2 cells. PRACTICAL APPLICATION: This work may suggest that FVP could be used as an adjuvant therapy to anticancer drugs, providing new application prospects and possibilities.

Auricularia polytricha and Flammulina velutipes reduce liver injury in DSS-induced Inflammatory Bowel Disease by improving inflammation, oxidative stress, and apoptosis through the regulation of TLR4/NF-κB signaling pathways.

Auricularia polytricha and Flammulina velutipes are two dietary mushrooms mostly consumed in China and known for their traditional use on gastric ulceration and to boost bowel movement. Considering the gut-liver axis, which has been recognized for its role in the autoimmune modulation, and the implications of the intestinal barrier in the pathogenesis of liver diseases that remain unclear, the therapeutic effects of A. polytricha (APE) and F. velutipes (FVE) on inflammatory bowel disease (IBD)-induced liver injury in mice was investigated as well as their potential mechanism via the signaling pathways they could involve. 3% DSS was administered to the mice in drinking water, to induce ulcerative colitis, followed by oral administration of APE and FVE. The biochemical, oxidative stress and inflammatory parameters, mRNA and protein expressions were assessed. The results revealed that DSS-induced liver histopathological changes were ameliorated by APE and FVE treatment. APE and FVE administration also improved the ALT and AST activity as well as the pro-inflammatory cytokines and oxidative factors. Data also showed that, in addition to their regulation of tight junctions' disruption, APE and FVE attenuated genes and proteins expression involved in apoptosis, lipid metabolism, and bile acid homeostasis via inhibiting TLR4/NF-κB and caspase signaling pathways and stimulating Keap1/Nrf2 signaling pathways. In conclusion, APE and FVE regulated liver injury on DSS-induced ulcerative colitis by alleviating inflammation, oxidative stress, and apoptosis, suggesting that they could be used as therapeutic alternatives against liver diseases in addition to their functions as dietary supplements.

Flammulina velutipes stem regulates oxidative damage and synthesis of yolk precursors in aging laying hens by regulating the liver-blood-ovary axis.

Egg production levels in late laying hens are negatively correlated with increasing age. Decreased liver and ovarian function in aging laying hens is accompanied by decreased antioxidant capacity, reproductive hormone levels, and follicular development, resulting in decreased synthesis of yolk precursors. The golden needle mushroom (Flammulina velutipes) has been reported to exhibit anti-inflammatory, antioxidant, and hypolipidemic properties. We aimed to reveal the therapeutic effects of F. velutipes stem (FVS) on liver-blood-ovary axis and investigate the underlying mechanisms. A total of 360 sixty-seven-wk-old laying hens were randomized into 4 treatment groups: 1) basal maize-soybean meal diet (CON); 2) basal maize + 20 g/kg FVS (2% FVS); 3) basal maize + 40 g/kg FVS (4% FVS); and 4) basal maize + 60 g/kg FVS (6% FVS). FVS groups demonstrated significantly increased egg production and ovarian development compared with the CON group. The addition of FVS increased the levels of antioxidant enzymes (GSH-Px, T-SOD, and T-AOC) in the liver, serum, and ovaries and decreased malondialdehyde levels by regulating the expression of proteins related to the Keap1-Nrf2/ARE signaling pathway. Additionally, FVS significantly decreased ovarian apoptosis by regulating Bax, Bcl-2, and caspase3 mRNA and protein expression levels. FVS significantly increased the expression levels of estradiol, progesterone, luteinizing hormone, and follicle stimulating hormone and their respective receptors. With increased levels of estradiol transported to the liver through the bloodstream, targeted binding to estrogen receptor (ER)-α and ER-ß led to significant increases in ApoVLDL II, ApoB, and VTG II mRNA expression associated with yolk precursor synthesis. FVS decreased the levels of triglyceride and total cholesterol and significantly increased the expression of lipid metabolism, and transport-related mRNAs (FAS, PPAR-a/γ, and MTTP) in the liver. Therefore, the dietary supplementation of FVS can maintain the productive performance of aging laying hens by alleviating the degree of oxidative stress and regulating the transport of functional substances along the liver-blood-ovary axis, thereby improving the synthesis of yolk precursors.

Flammulina velutipes Mycorrhizae Attenuate High Fat Diet-Induced Lipid Disorder, Oxidative Stress and Inflammation in the Liver and Perirenal Adipose Tissue of Mice.

Flammulina velutipes (FV) is edible mushroom that has nutritional and medicinal values. FV mycorrhizae, the by-products of FV, are an abundant source and receive less attention. The objective of this study was to investigate the composition of FV mycorrhizae, and its effects on high fat diet (HFD)-induced lipid disorder, oxidative stress, and inflammatory cytokines, both in the liver and perirenal adipose tissue (PAT) of mice. The results showed that FV mycorrhizae contain abundant trace elements, polysaccharide, amino acids and derivatives, and organic compounds. It was found that 4% FV mycorrhizae (HFDFV) supplementation decreased HFD-induced liver weight and triglyceride (TG) in the plasma, liver and PAT, altered plasma and hepatic fatty acids profiles, promoted gene expression involved in lipid hydrolysis, fatty acid transportation and ß-oxidation in the liver and reduced lipid synthesis in the liver and PAT. HFDFV attenuated HFD-induced oxidative stress and pro-inflammatory cytokine by increasing GSH/GSSG, and decreasing levels of MDA and IL6 both in the liver and PAT, while it differentially regulated gene expression of IL1ß, IL6, and CCL2 in liver and PAT. The results indicated that FV mycorrhizae are effective to attenuate HFD-induced lipid disorder, oxidative stress and inflammation in the liver and PAT, indicating their promising constituents for functional foods and herbal medicine.

Storage Quality Variation of Mushrooms (Flammulina velutipes) after Cold Plasma Treatment.

Flammulina velutipes is susceptible to mechanical damage, water loss, microbial growth, and other factors that lead to postharvest deterioration, thereby shortening the storage period. The purpose of this study was to analyze the effects of cold plasma treatment on the physicochemical properties and antioxidant capacity of F. velutipes during storage at 4 °C for 21 days. Compared to the control group, cold plasma cold sterilization (CPCS) treatment (150 Hz, 95 kV for 150 s) effectively inhibited the growth and multiplication of microorganisms on the surface of F. velutipes, with no significant effect on the fresh weight change and the superoxide anion generation rate, but with a higher postharvest 1,1-dephenyl-2-picrylhydrzyl (DPPH) clearance rate. Moreover, CPCS increased antioxidant enzyme activities, delayed both malondialdehyde (MDA) accumulation and vitamin C loss, inhibited the browning reaction and polyphenol oxidases (PPO) activity and protected F. velutipes cell membrane from disruption. In general, CPCS not only achieved bacteriostatic effects on F. velutipes during storage, but also reduced cell damage from free radical oxidation, resulting in better postharvest quality and longer shelf life.

The biological regulatory activities of Flammulina velutipes polysaccharide in mice intestinal microbiota, immune repertoire and heart transcriptome.

The effects of a novel Flammulina velutipes polysaccharide (FVP) on intestinal microbiota, immune repertoire and heart transcriptome were investigated in this study. The results showed that FVP treatment could effectively regulate the abundance of colonic microbiota. And FVP exhibited obvious immunoregulatory effect by influencing V gene and J gene fragments usage on TCRα chain. The usage frequency of TRBV1, TRBJ1-6 and TRBJ1-5 were significantly altered, and 41 V-J pairs were identified with obvious difference after FVP treatment. Furthermore, the mRNA of mice heart was analyzed by transcriptome assay. Total 525 genes and 1587 mRNA were significantly changed after FVP treatment. KEGG annotation indicated that the up-regulated mRNA was enriched in 17 pathways including adherens junction, mTOR signaling pathway, insulin signaling pathway, mitophagy, tight junction, PPAR signaling pathway and TNF signaling pathway, etc. Meanwhile, the down-regulated mRNA was gathered in AMPK signaling pathway, metabolism of xenobiotics by cytochrome P450, apelin signaling pathway, PPAR signaling pathway, PI3K-Akt signaling pathway, insulin signaling pathway, cardiac muscle contraction, adrenergic signaling in cardiomyocytes, Fc gamma R-mediated phagocytosis, etc. The great potential exhibited by FVP could make it an ideal candidate as complementary medicine or functional food for promotion of health.

Flammulina velutipes polysaccharide improves C57BL/6 mice gut health through regulation of intestine microbial metabolic activity.

Flammulina velutipes polysaccharides (FVP) can improve gut health through gut microbiota and metabolism regulation. In this study, the 28-days fed experiment was used to investigate gut microbime and metabolic profiling induced by FVP. After treatment, intestinal tissue section showed the higher villus height and villus height/crypt depth (V/C) value in FVP-treated group. The 16 s rRNA gene sequencing revealed microbiota composition alteration caused by FVP, as the Firmicutes phylum increased while Bacteroidetes phylum slightly decreased. The metabolic profiling was detected by LC/MS and results showed 56 and 99 compounds were dramatically changed after FVP treatment in positive and negative ion mode, respectively. Annotation in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways displayed the adjustment of energy metabolism, amino acid metabolism, nucleotide metabolism and other related basic pathways after FVP treatment. Our study suggested that FVP can be developed as a dietary supplement for intestine health promotion.

Biotechnological Production and Sensory Evaluation of ω1-Unsaturated Aldehydes.

Lipid extracts of the fungus Flammulina velutipes were found to contain various scarce fatty acids including dodec-11-enoic acid and di- and tri-unsaturated C16 isomers. A biotechnological approach using a heterologously expressed carboxylic acid reductase was developed to transform the fatty acids into the respective aldehydes, yielding inter alia dodec-11-enal. Supplementation studies gave insights into the fungal biosynthesis of this rarely occurring acid and suggested a terminal desaturation of lauric acid being responsible for its formation. A systematic structure-odor relationship assessment of terminally unsaturated aldehydes (C7-C13) revealed odor thresholds in the range of 0.24-22 µg/L in aqueous solution and 0.039-29 ng/L in air. In both cases, non-8-enal was identified as the most potent compound. All aldehydes exhibited green odor qualities. Short-chained substances were additionally associated with grassy, melon-, and cucumber-like notes, while longer-chained homologs smelled soapy and coriander leaf-like with partly herbaceous nuances. Dodec-11-enal turned out to be of highly pleasant scent without off-notes.

Dietary Supplementation with Flammulina velutipes Stem Waste on Growth Performance, Fecal Short Chain Fatty Acids and Serum Profile in Weaned Piglets.

This study was conducted to evaluate the effects of dietary FVS supplementation on growth performance, nutrient digestibility, biochemical profile of serum and fecal short chain fatty acids (SCFAs) production in weaned piglets. In Exp.1, 150 weaned pigs (initial body weight: 6.89 ± 1.17 kg) were allotted to five dietary treatments. The treatment diets included a basal diet and four experimental diets supplemented with 2.5%, 5.0%, 7.5% and 10.0% FVS respectively. The animal trial lasted for 28 days. In Exp.2, 72 piglets (initial body weight: 8.20 ± 1.67 kg) were allotted to three dietary treatments. The treatment diets included a basal diet and two experimental diets supplemented with 1.5% and 3.0% FVS, respectively. The animal trial lasted for 56 days. The results showed that pigs fed dietary FVS with 3% or lower inclusion levels had no significant difference (p > 0.10) on growth performance compared with pigs fed the control diet during day 1-28 and day 1-56. Dietary FVS supplementation decreased the apparent total tract digestibility (ATTD) of nutrients on day 28, day 35 and day 56, but no significant changes (p > 0.05) of nutrient digestibility were observed on day 14. Although piglets fed diets with higher levels of FVS showed impaired growth performance and ATTD of nutrients, dietary FVS supplementation improved the fecal SCFA production, antioxidant capacity, interleukin-2 and growth hormone levels in serum, and reduced the harmful low-density lipoprotein levels in serum on day 56. In conclusion, as a promising alternative fibrous ingredient, FVS could be supplemented in diets of weaned piglets with a proportion under 3%.

O/W Pickering emulsions stabilized by Flammulina velutipes polysaccharide nanoparticles as a fat substitute: the effects of phase separation on emulsified sausage's techno-functional and sensory quality.

BACKGROUND: The application of Pickering emulsion stabilized by food-derived particles is of great interest in the food field, including meat processing. However, the creaming phenomenon is a thorny problem and may impact the resulting product quality. Here, we used polysaccharide nanoparticles from Flammulina velutipes (FVPN) as a stabilizer to prepare a oil/water Pickering emulsion and partly replace the original fat of common emulsified sausage, focusing on exploring the influence of phase separation on the sausage's techno-functional and sensory quality, with the aim of developing a new alternative fat substitute. RESULTS: Reformulated sausages showed increases in moisture (53.24-64.85%) and protein content (11.97-12.76%), but were reduced in fat content (27.28-18.76%). The increased FPOE (FVPN-palm oil emulsion; substitution rate 5-37%) amount in sausages resulted in significantly reduced (P < 0.05) cooking loss (18.87-8.63%). Meanwhile, emulsion improved the springiness and cohesiveness of sausage and significantly reduced (P < 0.05) hardness and chewiness when the replacement amount was less than 20%. Experimental sausages attained a more compact pore structure without harming sensory characteristics. Compared with creaming emulsion, pristine emulsion resulted in a sausage with higher moisture content, lower cooking loss, better elasticity and denser structure. CONCLUSION: The characteristics of sausages could be influenced by emulsion stability. Emulsion, especially with no creaming, can be effectively used as fat substitute at a level of 20% or less without adversely affecting the sensory characteristics of emulsified sausages. The incorporation of FPOE provides the potential for developing a new alternative approach for animal fat improvement in meat products. © 2019 Society of Chemical Industry.

Overexpression of the saccharopine dehydrogenase gene improves lysine biosynthesis in Flammulina velutipes.

Saccharopine dehydrogenase (EC 1.5.1.7) regulates the last step of fungal lysine biosynthesis. The gene (Fvsdh) encoding saccharopine dehydrogenase was identified and cloned from the whole genome of Flammulina velutipes. The genomic DNA of Fvsdh is 1257 bp, comprising three introns and four exons. The full-length complementary DNA of Fvsdh comprises 1107 bp with a deduced amino acid sequence of 368 residues. A 1,000-bp promoter sequence containing the TATA box, CAAT box, and several putative cis-acting elements was also identified. The results of tissue expression analysis showed that the expression level of the Fvsdh gene was higher in the pileus than in the stipe whether in the elongation or maturation stage. Further research showed that the lysine contents were 3.03 and 2.95 mg/g in maturation-pileus and elongation-pileus, respectively. In contrast, the lysine contents were 2.49 and 2.07 mg/g in elongation-stipe and maturation-stipe, respectively. To study the function of Fvsdh, we overexpressed Fvsdh in F. velutipes and found that Fvsdh gene expression was increased from 1.1- to 3-fold in randomly selected transgenic strains. The lysine contents were also increased from 1.12- to 1.3-fold in these five transformants, except for strain T3, in which the lysine contents were the same as the control. These results indicate that the expression of the Fvsdh gene can affect the lysine content of F. velutipes.

Combination of on-line desalting and HPLC-UV-ESI-MS for simultaneous detection and identification of FIP-fve and flammutoxin in Flammulina velutipes.

A rapid analytical approach, on-line desalting HPLC-UV-ESI-MS method, for the analysis of FIP-fve and flammutoxin (FTX), two important bioactive proteins in the fruiting bodies of Flammulina velutipes, was developed. In this study, a highly efficient desalting method is provided using molecular weight cut-off centrifugal filtration and on-line desalting. Sample preparation followed by an on-line desalting HPLC-UV-ESI-MS system was employed for simultaneous desalting and detection and identification of FIP-fve and FTX. Results indicated that using trifluoroacetic acid as a modifier on a C18 reversed-phase column renders effective separation. ESI-MS revealed that the apparent molecular masses of FIP-fve and FTX were 12,749.1 Da and 21,912.5 Da, respectively. Eleven milligrams of FIP-fve was obtained from 100 g of fresh fruiting bodies, and UV detection was performed at 280 nm using bovine serum albumin as the standard protein. The calibration curve was linear in the concentration range of 0.29-4.69 mg/mL (r2 = 0.9999). FTX and a series of degradation products were isolated from F. velutipes using 35% saturated ammonium sulfate on a DEAE cellulose column. The complete identification of FTX and a series of degradation products were carried out by precipitation of various ammonium sulfate concentrations (0-45%, 45-65% and 65-90%), in-gel trypsin digestion, and MS analysis with combined database search. The molecular weights of FTX and a series of degradation products were 29,957.2 Da, 27,480.2 Da, 26,512.5 Da, and 21,912.5 Da.

Flammulina velutipes treatment of non-sterile tall wheat grass for enhancing biodegradability and methane production.

In this study fungal pretreatment of non-sterile tall wheat grass via the white rot fungi Flammulina velutipes was studied and the effect on biodegradability of lignocellulosic biomass and methane production, was evaluated. Degradation of lignin, cellulose, hemicellulose, and dry matter in non-sterile tall wheat grass during 28 days of fungal pretreatment using different inoculum ratio (0%-50%) and moisture content (MC) (45% MC, 65% MC, and 75% MC) were assessed via comparison to untreated biomass. Pretreatment with F. velutipes was most effective at 65% MC and 40% inoculum ratio, resulting in 22% lignin removal. The corresponding methane yields were 181.3 Ndm3·kg VS-1, which were 280% higher than for the untreated tall wheat grass.

Cognitive-enhancing effect of polysaccharides from Flammulina velutipes on Alzheimer's disease by compatibilizing with ginsenosides.

This study was performed to evaluate the cognitive-enhancing effect Flammulina velutipes polysaccharides (FVP) by compatibilizing with ginsenosides on D-galactose-induced Alzheimer's disease (AD) rats. Results of the Morris water maze test suggested that the cognitive ability of AD rats was significantly elevated after combined treatment with FVP and ginsenosides. Moreover, the anti-oxidant effect was also elevated as reflected by the increased levels of superoxide dismutase, catalase, and glutathione peroxidase and decreased levels of malondialdehyde in the combination group. Terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end labeling (TUNEL), immunohistochemistry, and Western blot analysis showed that the TUNEL apoptosis cell rate, Bax, cytochrome C, caspase-3, caspase-9, and apoptosis-inducing factor expression levels were obviously decreased, and the Bcl-2 expression levels were obviously increased in the hippocampi of rats treated with FVP and ginsenosides. The results suggested that the beneficial effect of FVP against AD can be significantly elevated by compatibilizing with ginsenosides. These findings provide scientific evidence supporting the use of FVP as a safe and effective drug to prevent and treat AD.

Improvement of Diet-induced Obesity by Ingestion of Mushroom Chitosan Prepared from Flammulina velutipes.

The anti-obesity effects of mushroom chitosan prepared from Flammulina velutipes were investigated using an animal model with diet-induced obesity. In this study, 5-week-old imprinting control region (ICR) mice were divided into six groups of 10 mice each and fed different diets based on the MF powdered diet (standard diet) for 6 weeks: standard diet control group, high-fat diet control group (induced dietary obesity) consisting of the standard diet and 20% lard, and mushroom chitosan groups consisting of the high-fat diet with mushroom chitosan added at 100, 500, 1,000, and 2,000 mg/kg body weight. On the final day of the experiment, mean body weight was 39.1 g in the high-fat control group and 36.3 g in the 2,000 mg/kg mushroom chitosan group, compared to 35.8 g in the standard diet control group. In the mushroom chitosan groups, a dose-dependent suppression of weight gain and marked improvements in serum triglycerides, total cholesterol, LDL-cholesterol, and HDL-cholesterol were found. The mushroom chitosan groups showed fewer and smaller fat deposits in liver cells than the high-fat diet control group, and liver weight was significantly reduced. Glutamic oxaloacetic transaminase (GOT) and glutamate pyruvic transaminase (GPT), which are indices of the hepatic function, all showed dose-dependent improvement with mushroom chitosan administration. These results suggested that mushroom chitosan acts to suppress enlargement of the liver from fat deposition resulting from a high-fat diet and to restore hepatic function. The lipid content of feces showed a marked increase correlated with the mushroom chitosan dose. These findings suggest the potential use of mushroom chitosan as a functional food ingredient that contributes to the prevention or improvement of dietary obesity by inhibiting digestion and absorption of fats in the digestive tract and simultaneously promotes lipolysis in adipocytes.

Oral fungal immunomodulatory protein-Flammulina velutipes has influence on pulmonary inflammatory process and potential treatment for allergic airway disease: A mouse model.

BACKGROUND/PURPOSE: House dust mite (HDM) is well known as one of the major indoor allergens that trigger allergic inflammation, especially asthma, and accounts for 85% of all cases. So far, asthma has been thought of as a condition of imbalance between T helper (Th)1 and Th2. Fungal immunomodulatory protein-Flammulina velutipes (FIP-fve) has been seemingly demonstrated to modulate the response to Th1 cytokine production. The aim of this study was to investigate if the oral administration of FIP-fve can inhibit HDM-induced asthma inflammation in the mouse model. METHODS: We divided the mice (female BALB/c, 4-6 weeks) into four groups: the prevention group, which consisted of mice sensitized by HDM (intraperitoneally on Day 1, Day 7, and Day 14, and intranasally on Day 14, Day 17, Day 21, Day 24, and Day 27) fed with FIP-fve from Day 1 to Day 14; the treatment group, which comprised mice that received treatment from Day 14 to Day 28; the positive control (PC, sensitized by HDM fed without FIP-fve) group; and the negative control group (NC, nonsensitized). Airway hyperresponsiveness induced by methacholine challenge was determined using whole-body barometric plethysmography. In addition, cytokines were analyzed from bronchoalveolar lavage fluid and serum. Histopathological studies and Liu's staining method in mice lungs were also performed. RESULTS: The results showed that both pre- and posttreated FIP-fve groups had significantly reduced airway hyperresponsiveness compared with the PC group after methacholine challenge. In addition, a significantly decreased level of HDM-specific immunoglobulin E in serum and decreased production of Th2 cytokines in bronchoalveolar lavage fluid and serum were observed in these two FIP-fve fed groups. Moreover, more decreased amounts of infiltrating inflammatory cells were present in the lungs of FIP-fve fed groups than those of the PC group. CONCLUSION: Oral FIP-fve had an anti-inflammatory effect on the acute phase of the airway inflammatory process induced by HDM in the mouse model and might have a potentially therapeutic role for allergic airway diseases.

Selenium uptake, tolerance and reduction in Flammulina velutipes supplied with selenite.

Recently, selenium (Se) enriched mushrooms have been exploited as dietary Se supplements, but our knowledge of the metabolic process during the Se enrichment process is far from complete. In this study, the uptake, tolerance and reduction of selenite in a widely cultivated mushroom, Flammulina velutipes, was investigated. The results showed that pH variation (from 5.5-7.5), metabolic inhibitor (0.1 mM 2,4-DNP) and P or S starvation led to 11-26% decreases in the selenite uptake rate of F. velutipes. This indicates that a minor portion of the selenite uptake was metabolism dependent, whereas a carrier-facilitated passive transport may be crucial. Growth inhibition of F. velutipes initiated at 0.1 mM selenite (11% decrease in the growth rate) and complete growth inhibition occurred at 3 mM selenite. A selenite concentration of 0.03-0.1 mM was recommended to maintain the balance between mycelium production and Se enrichment. F. velutipes was capable of reducing selenite to elemental Se [Se(0)] including Se(0) nanoparticles, possibly as a detoxification mechanism. This process depended on both selenite concentration and metabolism activity. Overall, the data obtained provided some basic information for the cultivation of the selenized F. velutipes, and highlighted the opportunity of using mushrooms for the production of Se(0) nanoparticles.

Identification of flavonoids from Flammulina velutipes and its neuroprotective effect on pheochromocytoma-12 cells.

The flavonoids from Flammulina velutipes (FVF) were isolated and the chemical composition of FVF was identified by high-performance liquid chromatography/mass spectrometry (HPLC/MS). The antioxidant activity and the neuroprotective effect against H2O2-induced pheochromocytoma-12 cells (PC12) cytotoxicity of FVF was investigated. Six flavonoids from FVF were identified as arbutin, epicatechin, phillyrin, apigenin, kaempferol and formononetin. Obtained results suggested that FVF possesses strong antioxidant properties in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities as well as reducing power. Pretreatment of PC12 cells with FVF significantly increased the survival rate of cells, glutathione level and superoxide dismutase activity. Conversely, the pretreatment reduced the release of lactate dehydrogenase and the accumulation of both intracellular reactive oxygen species and malondialdehyde. These findings collectively suggested that the protective effects of FVF against H2O2-induced neurotoxicity in PC12 cells as well as the potential for neuroprotection of FVF could be due to its antioxidant action.

Cloning and Expression Analysis of Phenylalanine Ammonia-Lyase Gene in the Mycelium and Fruit Body of the Edible Mushroom Flammulina velutipes

Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi.

Multiple-fingerprint analysis for investigating quality control of Flammulina velutipes fruiting body polysaccharides.

Quality control issues overshadow potential health benefits of the edible mushroom Flammulina velutipes, with the detection and isolation of polysaccharides posing particular problems. In this study, multiple-fingerprint analysis was performed using chemometrics to assess polysaccharide quality and antioxidant activity of F. velutipes fruiting bodies from different sources. The authentic source exhibited differences in both oxygen radical absorbance capacity and ferric reducing antioxidant power from foreign sources. IR spectroscopic/HPLC fingerprints of polysaccharide extracts from the authentic source were established and applied to assess the polysaccharide quality of foreign sources. Analysis of IR fingerprints using Pearson correlation coefficient gave correlation coefficient r values of 0.788 and 0.828 for two foreign sources, respectively, indicating distinctness from the authentic source. Analysis of HPLC fingerprints using the supervised method by Traditional Chinese Medicine could not discriminate between sources (r > 0.9), but principal component analysis of IR and HPLC fingerprints distinguished the foreign sources.