Pholiota Adiposa Research Articles

First report on isolation of methyl gallate with antioxidant, anti-HIV-1 and HIV-1 enzyme inhibitory activities from a mushroom (Pholiota adiposa)

In this study, a compound with antioxidant and anti-HIV activities designated as HEB was first isolated from the edible mushroom Pholiota adiposa by extraction with ethanol and ethyl acetate. HEB was then purified by high performance liquid chromatography (HPLC) and identified to be methyl gallate (C8H8O5, 184.1Da) based on data from its mass spectrum (MS) and nuclear magnetic resonance (NMR) spectrum. HEB displayed strong antioxidant potency in inhibiting, at 1.36mM concentration, erythrocyte hemolysis and scavenging DPPH radicals and superoxide anion (O2−) by 82.4%, 85.6% and 71.4%, respectively. Besides exhibiting a low cytotoxicity, compound HEB demonstrated significant anti-HIV activity in that it inhibited HIV-1 replication in TZM-BL cells infected by pseudovirus with an IC50 value of 11.9μM. Further study disclosed that HEB inhibited the viral entry process and activities of key enzymes essential for the HIV-1 life cycle. HEB inhibited HIV-1 reverse transcriptase and integrase activities with an IC50 value of 80.1μM and 228.5μM, respectively, and at 10mM concentration inhibited HIV-1 protease activity by 17.1% which was higher than that achieved by the positive control pepstatin A. Interestingly, this study first revealed that H2O2 stimulation not only activated cell oxidative stress responses, but also accelerated HIV-1 long terminal repeat (LTR) promotion in TZM-BL cells, which was significantly reduced by HEB from 18.2% to about 2%. It implied a direct relationship between the antioxidant and anti-HIV activities of the natural active constituent HEB. Nuclear transcription factor kappa B (NF-κB) signal pathways plays an important role in oxidative stress responses. Meanwhile, there is κB target sequence in HIV promoter LTR which is significant for virus replication and gene expression. In this study, Western Blot assay showed that HEB could inhibit the activation of NF-κB signal pathway stimulated by H2O2 in mouse spleen cells through suppressing NF-κB (p65) translocation into nucleus and NF-kappa-B inhibitor (IκB) degradation in cytoplasm. In summary, the antioxidant HEB from P. adiposa could inhibit HIV-1 replication through multiple target sites. The data suggest that natural antioxidant compounds might have a potential for treatment of AIDS.

검은비늘버섯 분말을 첨가한 쿠키의 품질특성 및 항산화활성

Pholiota adiposa is an edible as well as medicinal mushroom cultured in China and Japan with anti-tumor, anti-microbial, anti-hypertensive, and anti-hyperlipidemic activities. This study was carried out to investigate the quality characteristics of cookies added with Pholiota adiposa powder. Chemical composition of Pholiota adipose powder was as follows: 3.8% moisture, 6.9% ash, 1% crude lipids, 3% protein, 53.3% carbohydrates, and 31.9% crude dietary fiber. To analyze quality characteristics during the manufacture of cookies, pH, texture, color, antioxidant activity, total polyphenol content, and sensory evaluation were determined. As the content of Pholiota adiposa powder increased, the pH of cookies decreased from 6.7 to 6.02. Hardness of cookies decreased with increasing Pholiota adiposa powder content, and cookies added with 5% powder were softer than the control. Regarding surface color, L-value decreased with increasing powder content compared to that of the control, whereas in surface color among various powder levels was not significantly different. As the content of Pholiota adiposa powder increased, both antioxidant activity and total polyphenol content increased. In the sensory evaluation, cookies added with 3% Pholiota adiposa powder showed remarkably higher values compared to control and other amples in all aspects.

Characterization of a β-1,4-glucosidase from a newly isolated strain of Pholiota adiposa and its application to the hydrolysis of biomass

The highly efficient β-1,4-glucosidase (BGL)-secreting strain, Pholiota adiposa SKU0714, was isolated and identified based on its morphological features and sequence analysis of internal transcribed spacer (ITS) rDNA. P. adiposa BGL (PaBGL), which contained a carbohydrate moiety, was purified to homogeneity from P. adiposa culture supernatants by 2-step chromatography on DEAE and Sephacryl gel filtration columns. The relative molecular weight of PaBGL was 60 kDa by SDS-PAGE or 59 kDa by size exclusion chromatography, indicating that the enzyme is a monomer. The pH and temperature optima for hydrolysis were 5.0 and 65 °C, respectively. PaBGL showed the highest activity towards p-nitrophenyl-β-d-glucopyranoside (Vmax = 4390 U mg protein−1, Km = 2.23 mol m−3) and cellobiose (Vmax = 3460 U mg protein−1, Km = 5.60 mol m−3) ever reported. Its internal amino acid sequences showed homology with hydrolases from the glycoside hydrolase family 3 (GH3), indicating that PaBGL is a member of the GH3 family. The hydrolysis of rice straw using a commercial cellulase, Celluclast® 1.5L, resulted in a higher saccharification yield with the addition of PaBGL than with Novozyme 188. PaBGL may be a good candidate for applications that convert biomasses to biofuels and chemicals.

Effects of Adenosine Extract from Pholiota adiposa (Fr.) Quel on mRNA Expressions of Superoxide Dismutase and Immunomodulatory Cytokines

Pholiota adiposa is a kind of edible mushroom which has long been known for its health care applications. To reveal the exact mechanism of its protective functions in humans, in this study we isolated and identified the active compound PB3 of P. adiposa for the first time by a combination of chromatography techniques, including NKA macroporous resin and Sephadex G-15. PB3, with molecular mass of 267.2 Da and molecular formula of C10H13N5O4 discovered by mass spectrum (MS) was identified to be adenosine. Mice were injected intraperitoneally with purified fraction PB3. Seven days after injection, we found a 1.5-fold increase of IL10 at the mRNA level, while a down regulated expression of IL-2, IL-6 and IFN-γ to 49.0%, 56.9% and 73.4%, respectively, was detected in spleen by real-time quantitative PCR. What’s more, SOD expression level was significantly increased by 1.6-fold compared to control. Fraction PB3 displayed anti-inflammatory potency and heightened SOD activity on the transcriptional level, which could be considered of further pharmaceutical or medication value.

Purification, composition analysis and antioxidant activity of different polysaccharides from the fruiting bodies of Pholiota adiposa

Three water-soluble polysaccharide fractions (PAP1-1, PAP1-2 and PAP2-1) were isolated and purified from the fruiting bodies of Pholiota adiposa by diethylaminoethanol (DEAE) Sepharose fast flow column and superdex TM 200 column chromatography on an AKTA Purifier system. Results of high-performance size-exclusion chromatography (HPSEC) indicated that the average molecular weights of PAP1-1, PAP1-2 and PAP2-1 were 2.3 x 10 6 Da, 8.8 x 10 3 Da and 2.1 x 10 6 Da, respectively. Monosaccharide compositions analysis revealed that PAP1-1 and PAP2-1 were both mainly composed of glucose, and PAP1-2 was composed of rhamnose and glucose with a mole ratio of 1:3.61. The evaluation of antioxidant activity suggested that PAP1-1 with β-configuration glycosidic bond exhibited stronger scavenging activity of superoxide radical and hydroxyl radical than PAP1-2 and PAP2-1 contained both α- and β-configuration glycosidic bond, and should be explored as a novel potential antioxidant. Available data obtained with in vitro models suggested that antioxidant activity of polysaccharide may be related to the configuration of glycosidic bond. Key words : Pholiota adipose , polysaccharide, purification, antioxidant activity.

Saccharification of poplar biomass by using lignocellulases from Pholiota adiposa

A basidiomycetous fungus, identified as Pholiota adiposa SKU0714 on the basis of morphological and phylogenetic analyses, was found to secrete efficient lignocellulose degrading enzymes. The strain showed maximum endoglucanase, cellobiohydrolase and β-glucosidase activities of 26, 32 and 39U/mL, respectively and also secreted xylanase, laccase, mannanase, and lignin peroxidase with activities of 1680, 0.12, 65 and 0.41U/mL, respectively when grown with rice straw as a carbon source. Among the various plant biomasses tested for saccharification, poplar biomass produced the maximum amount of reducing sugar. Response surface methodology was used to optimize hydrolysis parameters. A maximum saccharification yield of 83.4% (667mg/g-substrate), the highest yield from any plant biomass, was obtained with Populus biomass after 24h of hydrolysis. P. adiposa was proven to be a good choice for the production of reducing sugars from cellulosic biomass.

The Biological Characteristics and its Sequence Analysis of <i>Pholiota adiposa</i>

In this paper, the biological characteristics of Pholiota adiposa were systematically studied. The results showed that the ideal temperature range for growth is from 20 °C to 25°C, with optimal temperature at 25°C; the optimal light condition is full darkness; the ideal pH range for growth is from 5 to 9, with optimal pH at 6; the preferred carbon source is sucrose, followed by glucose; the preferred nitrogen source is potassium nitrate, glutamic acid. The internal transcribed spacer region (ITS) was sequenced to determine whether the DNA sequence data supported the experimental result. The phylogenetic tree for the 19 pieces of homologous sequences were analyzed, with the highest homology reaching 99%.

Immobilization of Pholiota adiposa xylanase onto SiO2 nanoparticles and its application for production of xylooligosaccharides

Enhanced yields of different lignocellulases were obtained under statistically-optimized parameters using Pholiota adiposa. The k (cat) value (4,261 s(-1)) of purified xylanase under standard assay conditions was the highest value ever reported. On covalent immobilization of the crude xylanase preparation onto functionalized silicon oxide nanoparticles, 66 % of the loaded enzyme was retained on the particle. Immobilized enzyme gave 45 % higher concentrations of xylooligosaccharides compared to the free enzyme. After 17 cycles, the immobilized enzyme retained 97 % of the original activity, demonstrating its prospects for the synthesis of xylooligosaccharides in industrial applications.

Extraction and in vitro antioxidant activity of exopolysaccharide by Pholiota adiposa SX-01

The extraction parameters for Pholiota adiposa SX-01 exopolysaccharide (EPS) produced during submerged culture were optimized using response surface methodology. The optimum conditions for EPS extraction were predicted to be, concentration at 87.36°C, precipitation for 35.18 h at pH 8.81, and EPS production was estimated to be 8.45 g/L. The actual yield of EPS under these conditions was 8.39 g/L. The in vitro antioxidant activities of the EPS were investigated and results showed that the inhibition effects of EPS at a dosage of 250 mg/L on superoxide anion radical, hydroxyl radical and reducing power were 70.63 ± 5.26, 58.22 ± 5.41 and 0.37 ± 0.02% (absorbance at 700 nm), respectively. The results provide a reference for large-scale extraction of EPS by P. adiopsa SX-01 in industrial fermentation. Key words: Pholiota adiposa SX-01, exopolysaccharide, response surface methodology, antioxidant activity, in vitro

Isolation and Purification of Polysaccharides with Anti-tumor Activity from Pholiota adiposa (Batsch) P. Kumm. (Higher Basidiomycetes)

Pholiota adiposa is a mushroom with excellent medicinal and nutritional properties. After culture in fermentation medium, Ph. Adiposa mycelia were filtered, lyophilized, and powdered. A crude polysaccharide (PAP) of Ph. Adiposa was prepared from the mycelial powder with hot water, centrifuged, and the resulting supernatant lyophilized. PAP was fractionated by 30%, 60%, and 80% ethanol precipitation steps to yield PAP30, PAP60, and PAP80. Subsequently PAP30-1 and PAP30-2, PAP60-1 and PAP60-2, and PAP80-1 and PAP80-2 were isolated from PAP30, PAP60, and PAP80, respectively, by ion-exchange chromatography on a DEAE-Sepharose column. Polysaccharide content increased from 43.8% in PAP to 50.54%~73.19% in PAP30-1~PAP80-2. The protein content was 4.92% at minimum in these polysaccharide products. In order to identify the chemical composition, the six polysaccharides (PAP30-1, PAP30-2, PAP60-1, PAP60-2, PAP80-1, and PAP80-2) were further purified by gel filtration on Sephacryl S-100-500. Finally, three water-soluble polysaccharides (PAP30-2a, PAP60-2b, and PAP80-2a) were obtained. HPLC analysis revealed that PAP30-2a, PAP60-2b, and PAP80-2a exhibited a molecular weight of 6.6 × 105 Da, 8.4 × 103 Da, and 3.5 × 103 Da, respectively. The glucose content in PAP80-2a, PAP60-2b, and PAP30-2a was 57.8%, 72.7%, and 68.9%, respectively. PAP30-2a, PAP60-2b, and PAP80-2a demonstrated significant differences in anti-tumor activity in mice. PAP80-2a is the optimal bioactive constituent with anti-tumor and T-lymphocyte proliferation stimulating effects.

Extraction and in vitro antioxidant activity of intracellular polysaccharide by Pholiota adiposa SX-02

Response surface methodology (RSM) was used to optimize the extraction parameters for Pholiota adiposa SX-02 intracellular polysaccharide (IPS) produced during submerged culture. The optimum conditions of IPS extraction were predicted to be, ultrasonic power at 564.93 W, precipitation time 30.34 h and pH 8.28, and IPS yield was estimated at 19.75%. The actual value of IPS under these conditions was 20.51%. The in vitro antioxidant results showed that the inhibition effects of IPS at a dosage of 250 mg/l on superoxide anion, hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were 74.66 ± 5.31%, 69.20 ± 5.13%, and 75.20 ± 6.73%, respectively, which were 27.51 ± 2.23%, 16.58 ± 1.33%, and 9.46 ± 0.72% higher than that of butylated hydroxytoluene (BHT), respectively. The reducing power of IPS was 0.32 ± 0.02 (absorbance at 700 nm), 39.13 ± 3.47% higher than that of BHT. The results provide a reference for large-scale extraction of IPS by P. adiposa SX-02 in industrial fermentation and the IPS can be used as a potential antioxidant which enhances adaptive immune responses.

Biological characteristics of five wood-rotting fungi and wood-decaying ability to Betula platyphylla

In this paper, the growth rate of five wood-rotting fungi, i.e., Coriolus versicolor, Irpex lacteus, Fomes fomentarius, Piptoporus betulinus and Pholiota adiposa, in solid medium and their biomass in liquid culture medium were compared by measuring mycelium length and dry mass. The activity of three main ligninolytic enzymes in those fungi, namely LiP, MnP and Lac, were also tested by colorimetry. At the same time, these fungi were used to decay the wood samples from 300 natural trees of white birch, to study their wood-decaying ability by measuring wood mass loss. The result showed that the growth rate, biomass, ligninolytic enzyme activity, and wood-decaying ability of the fungi were incompletely correlated. The growth rates of C. versicolor and I. lacteus were faster than those of P. betulinus and F. fomentarius; P. adiposa was the slowest in growth. The biomass of P. betulinus was the highest; C. versicolor, I. lacteus and F. fomentarius were in the middle, and P. adiposa was the lowest. There existed LiP, MnP and Lac activities in all fungi except P. betulinus, and the enzyme activities induced by wood powder were all higher than those of the control. The Lac of I. lacteus and the LiP of F. fomentarius and P. adiposa were only expressed in wood powder medium; the longer the fungi were cultured, the higher activity the enzyme had. The decomposition ability of C. versicolor to wood samples was the highest, followed by F. fomentarius and P. betulinus; I. lacteus and P. adiposa were the lowest.

A novel lectin with antiproliferative activity from the medicinal mushroom Pholiota adiposa.

Little was known about biological activities of compounds from the medicinal mushroom of the genus Pholiota. A lectin from the Pholiota adiposa has now been isolated and its properties tested. The isolation procedure included ion exchange chromatography on DEAE-cellulose and CM-cellulose, and fast protein liquid chromatography-gel filtration (FPLC) on Superdex 75. The lectin was composed of two identical subunits, each with a molecular mass of 16 kDa. Its N-terminal amino-acid sequence showed little similarity to sequences of other Agaricales lectins. The hemagglutinating activity of the lectin was stable at temperatures up to 50 degrees C, and in NaOH and HCl solutions with concentrations less than 25 mM. It was inhibited by inulin (12.5-200 mM), but enhanced by Cu(2+) (6.25-25 mM), Fe(2+) (12.5-25 mM), and Al(3+) (6.25-25 mM) ions. The lectin showed antiproliferative activity toward hepatoma Hep G2 cells and breast cancer MCF7 cells with an IC(50) of 2.1 microM and approximately 3.2 microM, respectively. It exhibited HIV-1 reverse transcriptase inhibitory activity with an IC(50) of 1.9 microM. When compared with P. aurivella lectin, the only Pholiota lectin published to date, P. adiposa lectin differs in chromatographic behavior, molecular mass, N-terminal sequence, and effect of cations on hemagglutinating activity. In the case of the lectin from P. aurivella, its antifungal, antiproliferative, and HIV-1 reverse transcriptase inhibitory activities have not been determined.

A New Spiroaxane Sesquiterpene from Cultures of the Basidiomycete Pholiota adiposa

A new spiroaxane sesquiterpene (15-hydroxy-6α,12-epoxy-7β ,10αH,11β H-spiroax-4-ene, 1) was isolated from the culture broth of the basidiomycete Pholiota adiposa. The structure of 1 was established on the basis of MS, IR, ID, and 2D NMR experiments.

Characterization of a novel β-hydroxy-β-methyl glutaryl coenzyme A reductase-inhibitor from the mushroom,Pholiota adiposa

This study describes the extraction and characterization of a novel inhibitor against β-hydroxy-β-methyl glutaryl (HMG) Coenzyme A (CoA) reductase from the mushroomPholiota adiposa. Methanol extracts ofP. adiposa PAD-022 fruiting body showed the highest HMG-CoA reductase-inhibitory activity of 55.8%. The HMG-CoA reductase-inhibitor,P. adipos a PAD-022, was maximally extracted when its fruiting body was treated with methanol at 30°C for 12 h. The HMG-CoA reductase-inhibitor was obtained by systematic solvent extraction methods followed by gel column chromatography, and RP-HPLC. The purified product was found to possess an activity of IC50 6.8 μg and a yield of 1.8%. The molecular weight of purified HMG-CoA reductase-inhibitor was deduced to be 412.7 Da. The inhibitor was identified as stigmasterol (C29H46O) by serial instrumental analyses, including LC-Mass. NMR, FT-IR, and UV spectrometry.

Effect of a Pholiota adiposa Extract on Fat Mass in Hyperlipidemic Mice

The purpose of this study was to investigate the effect of a Pholiota adiposa extract on fat mass in hyperlipidemic mice fed on a high-fat diet. The water extracts from P. adiposa (ASI 24018) were not affected in the total triglyceride contents and epididymal fat mass in mice fed on a high-fat diet, but the retroperitoneal fat mass decreased significantly. This result suggests that the P. adiposa extract may be a potential candidate for use as a functional food that can act as a prophylactic against hyperlipidemia. However, the P. adiposa extract showed no effect in the total triglyceride contents and epididymal fat mass.

Effects of Lycii fructus and Edible Mushroom, Pholiota adiposa, on the Quality and Angiotensin I-Converting Enzyme Inhibitory Activity of Korean Traditional Rice Wine

The goal of this study was to develop a high value Korean traditional rice wine possessing the antihypertensive functionality. The effects of the Lycii fructus and Pholiota adiposa on the alcohol fermentation and its angiotensin I-converting (ACE) inhibitory activity of Korean traditional rice wine, yakju were investigated. Optimal fermentation conditions for the preparation of yakju consisted of the koji added at 10% and a fermentation period of 10 days at 25˚C. The best acceptability appeared with the Lf-1 traditional rice wine that was brewed by the addition of 1% Lycii fructus into the mash containing cooked rice, koji and antihypertensive S. cerevisiae. Its antihypertensive ACE inhibitory activity was 76%. Its ACE inhibitory activity was also increased about 8% by the addition of 0.1% of the edible mushroom Pholiota adiposa ASI 24012 fruiting bodies into the mash, showing good sensory characteristics. From these results, Lf-Pa1 yakju which was brewed by addition of 1% Lycii fructus and 0.1% Pholiota adiposa into the mash, has the potential to become a new functional Korean traditional rice wine with high antihypertensive properties.

비늘버섯으로부터 생리기능성 물질의 탐색

비늘버섯에 함유되어 있는 몇 가지 심혈관 질환과 노화예방 관련 생리 기능성 물질을 탐색하여 고부가가치의 건강 제품을 개발하고자 먼저 전국 각지에서 수집하고, 농업과학기술원, 인천대학 버섯 종균 은행 등으로부터 분양받은 비늘버섯 균주들을 인공 재배하여 63 균주의 자실체를 생산하였다. 이들 자실체들의 물과 메탄을 추출물을 각각 제조한 후 각종 생리 기능성을 조사 한 결과, 혈전용해 활성은 P. adiposa PAD-030 균주의 메탄을 추출물이 11.5 U, HMG-CoA reductase 저해활성은 P. adiposa PAD-022 균주의 메탄올 추출물이 76.8%로 가장 높았다. 항산화 활성과 tyrosinase 저해활성은 각각 Pholiota sp. PSP-015 균주의 물 추출물과 P. nameko PNA-024의 메탄올 추출물이 각각 72.7%, 69.5%로 우수하였다. 그러나 SOD 유사활성과 elastase 저해활성은 대부분의 비늘버섯 추출물에서 미약하거나 없었다. 위와 같이 조사된 몇 가지 생리 기능성 중 가장 우수한 활성을 보인 것은 P. adiposa PAD-022 균주 자실체의 메탄올 추출물의 HMG-CoA reductase 저해활성이었으며 이 저해 물질은 자실체 분말을 메탄올로 <TEX>$30^{\circ}C$</TEX>에서 12시간 진탕 시켰을 때 가장 많이 추출 되었다. Extracts from 63 kinds of Pholiota sp. fruiting bodies were prepared using water and methanol, and then their physiological functionalities were investigated. The methanol extracts from Pholiota adiposa PAD030 showed high fibrinolytic activity and those of P. adiposa ASI PAD-022 showed potential inhibitory activity of 76.8% against <TEX>${\beta}-hydroxy-{\beta}-methylglutaryl(HMG)-CoA$</TEX> reductase. The highest antioxidant and tyrosinase inhibitory activities were found in the water extracts of Pholiota sp. PSP-015 (72.7%) and methanol extracts of P. nameko PNA-024 (69.5%), respectively. However, superoxide dismutase(SOD)-like activity and elastase inhibitory activity were low in almost of the extracts. The HMG-CoA reductase inhibitor from the fruiting body of P. adiposa PAD-022 which showed the highest functionality was extracted maximally when powder of the fruiting body was shaked at <TEX>$30^{\circ}C$</TEX> for 12 h by methanol and its HMG-CoA reductase inhibitory activity was 80.2%.

Antimicrobial activity of the macrofungus Pholiota adiposa

The 60% methanolic extract of Pholiota adiposa exhibited antimicrobial activity.

Antimicrobial activity of the macrofungus Pholiota adiposa

The 60% methanolic extract of Pholiota adiposa exhibited antimicrobial activity.