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PubMed2004

A mushroom (Ganoderma capense) lectin with spectacular thermostability, potent mitogenic activity on splenocytes, and antiproliferative activity toward tumor cells.

An 18-kDa lectin, with an N-terminal sequence displaying slight similarity to some lectins and fungal immunomodulatory proteins, was isolated from the mushroom Ganoderma capense (Lloyd) Teng. It exhibited more potent mitogenic activity than that of concanavalin A toward mouse splenocytes, and antiproliferative activity toward leukemia (L1210 and M1) cells and hepatoma (HepG2) cells. The isolation procedure entailed ion exchange chromatography on Q-Sepharose, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 75. D(+)-galactose and D(+)-galactosamine specifically inhibited the hemagglutinating activity of the lectin. The hemagglutinating activity of the lectin was not affected over the temperature range 0-100 degrees C and after exposure to 100 degrees C for 60min. The activity was stable in the pH range of 4-11, and after incubation with solutions of various chlorides (from 3.125 to 50mM) including NaCl, KCl, CaCl(2), MgCl(2), ZnCl(2), MnCl(2), and AlCl(3). However, it was potentiated by 12.5-50mM FeCl(3). The lectin was devoid of HIV-1 reverse transcriptase inhibitory and antifungal activities.

Amino Acid Sequence↗
PubMed2004

Successful resonance Raman study of cresolase activity of mushroom tyrosinase.

Mono-oxygenase (cresolase) activity of mushroom tyrosinase (MT) in the presence of 4-[(4-hydroxyphenyl)azo]-benzenesulfonamide (HPABS) was successfully studied by resonance Raman (rR) spectroscopy. HPABS is a synthetic competitive inhibitor (K(i)=7.17 x 10(-6)M) for the cresolase activity with a large extinction coefficient at 365 nm. Upon reacting with MT, HPABS produced an enzyme-inhibitor (EI) complex with sufficiently long life span. Analyzing the ensuing spectrum indicates that the azo tautomer of HPABS binds to the enzyme and retains its geometrical isomeric form in the EI complex. The observed changes in the rR spectrum of HPABS after binding to MT support the idea that an electrophilic attack on the inhibitor has happened. Similar experiments were designed for studying the oxidase activity of MT. However, the enzymatic reaction, even in the presence of 4-[(2,4-dinitrophenyl)azo]-1,2-benzenediols was still fast enough to tan the reaction solution quickly and render its rR spectrum impregnable background.

Agaricales↗
PubMed2004

Purification of a novel low-molecular-mass laccase with HIV-1 reverse transcriptase inhibitory activity from the mushroom Tricholoma giganteum.

A laccase with a novel N-terminal sequence, a low molecular mass of 43 kDa smaller than those of previously reported laccases, a pH optimum of 4, and a temperature optimum at 70 degrees C was isolated from fresh fruiting bodies of the mushroom Tricholoma giganteum. The activity of the enzyme rose steadily from 20 to 50 degrees C, increased very slowly from 50 to 70 degrees C, and fell slightly when the temperature was further increased to 80 degrees C. The activity of the laccase underwent little changes over the pH range 3.0-5.0. However, the enzyme activity dwindled to nothing after exposure to 100 degrees C for 10 min and when the ambient pH was 7 or above. The procedure used for purifying the enzyme included ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose, and FPLC-gel filtration on Superdex 75. The laccase was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-cellulose. It inhibited HIV-1 reverse transcriptase with an IC(50) of 2.2 microM.

Agaricales↗
PubMed2004

A new laccase from dried fruiting bodies of the monkey head mushroom Hericium erinaceum.

A laccase with a novel N-terminal sequence, a molecular mass of 63kDa, and inhibitory activity toward HIV-1 reverse transcriptase (IC(50)=9.5microM) was isolated from dried fruiting bodies of the monkey head mushroom Hericium erinaceum. A chromatographic procedure involving ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose and fast protein liquid chromatography-gel filtration on Superdex 75 was employed. The laccase was adsorbed on DEAE-cellulose and Q-Sepharose but unadsorbed on CM-cellulose. High activity of the enzyme was observed at pH 3-5 and at 50-80 degrees C. Its activity was completely abolished at pH 8 and 9 and after boiling for 10min. A temperature of 50 degrees C and a pH of 5.0 were optimal for its activity.

Agaricales↗
PubMed2005

A novel homodimeric lactose-binding lectin from the edible split gill medicinal mushroom Schizophyllum commune.

A homodimeric lactose-binding lectin with a molecular mass of 64kDa was isolated from fresh fruiting bodies of the split gill mushroom Schizophyllum commune. The N-terminal sequence of the lectin is similar to a part of the sequence of the cell division protein from Gleobacter violaceus. The lectin was isolated by using a procedure which involved ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of the lectin was stable at temperatures up to 40 degrees C, and in concentrations of NaOH and HCl solution up to 125 and 25mM, respectively. The lectin exhibited potent mitogenic activity toward mouse splenocytes, antiproliferative activity toward tumor cell lines, and inhibitory activity toward HIV-1 reverse transcriptase. It was devoid of antifungal activity.

Amino Acid Sequence↗
PubMed2006

Isolation and characterization of a novel lectin from the mushroom Armillaria luteo-virens.

From the dried fruiting bodies of the mushroom Armillaria luteo-virens, a dimeric lectin with a molecular mass of 29.4 kDa has been isolated. The purification procedure involved (NH(4))(2)SO(4) precipitation, ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of the lectin could not be inhibited by simple sugars but was inhibited by the polysaccharide inulin. The activity was stable up to 70 degrees C but was acid- and alkali-labile. Salts including FeCl(3), AlCl(3), and ZnCl(2) inhibited the activity whereas MgCl(2), MnCl(2), and CaCl(2) did not. The lectin stimulated mitogenic response of mouse splenocytes with the maximal response achieved by 1microM lectin. Proliferation of tumor cells including MBL2 cells, HeLa cells, and L1210 cells was inhibited by the lectin with an IC(50) of 2.5, 5, and 10 microM, respectively. However, proliferation of HepG2 cells was not affected. The novel aspects of the isolated lectin include a novel N-terminal sequence, fair thermostability, acid stability, and alkali stability, together with potent mitogenic activity toward spleen cells and antiproliferative activity toward tumor cells.

Agaricales↗
PubMed2004

Oxidation products of quercetin catalyzed by mushroom tyrosinase.

Quercetin was oxidized as a substrate catalyzed by mushroom tyrosinase to the corresponding o-quinone and subsequent isomerization to p-quinone methide type intermediate; followed by the addition of water on C-2 yielding a relatively stable intermediate, 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone. In the presence of a catalytic amount of l-DOPA as a cofactor, the rate of this oxidation was enhanced. Fisetin, which lacks the C-5 hydroxyl group, was also oxidized but the rate of oxidation was faster than that of quercetin, indicating that the C-5 hydroxyl group is not essential but is associated with the activity.

Agaricales↗
PubMed2006

Inhibitory effects of phloridzin dihydrate on the activity of mushroom (Agaricus bisporus) tyrosinase.

The inhibitory effects of phloridzin dihydrate on the activity of mushroom tyrosinase have been studied. The results show that phloridzin can inhibit the diphenolase activity of the enzyme and the inhibition displays to be reversible. The IC(50) value was estimated as 110microM. The kinetic analysis showed that the inhibition of phloridzin on the diphenolase activity of the enzyme is of competitive type, and the inhibition constant (K(I)) was determined to be 64.3microM. The inhibitory effects of the different concentrations of phloridzin on the monophenolase activity were also studied. There were almost no changes in the lag period and the steady-state rate, while the plateaus in the inhibitory curve lowered with increasing the concentration of phloridzin when using tyrosine as a substrate.

Agaricales↗
PubMed2006

An aqueous polysaccharide extract from the edible mushroom Pleurotus ostreatus induces anti-proliferative and pro-apoptotic effects on HT-29 colon cancer cells.

Anti-proliferative and pro-apoptotic activities of fractions of Pleurotus ostreatus were examined using HT-29 colon cancer cells in vitro. A hot-water-soluble fraction of the mycelium of the liquid cultured mushroom was partially isolated and chemically characterized as a low-molecular-weight alpha-glucan. HT-29 cells were exposed to the different isolates and significant inhibition of proliferation was obtained in a dose-dependent manner. Proliferation inhibition was shown to be the result of apoptotic induction because the pro-apoptotic molecules Bax and cytosolic cytochrome-c were upregulated. Fluorescence-activated cell sorter analyses of polysaccharide-treated HT-29 cells showed a high percentage of Annexin-positive cells. Here, we describe a newly identified low-molecular-weight alpha-glucan with promising anti-tumorigenic properties, and demonstrate its direct effect on colon cancer cell proliferation via induction of programmed cell death.

Annexin A5↗
PubMed2004

Structural studies of water-soluble polysaccharides of an edible mushroom, Termitomyces eurhizus. A reinvestigation.

The structure of two polysaccharides isolated from the hot aqueous extract of fruiting bodies of the mushroom, Termitomyces eurhizus, have been reinvestigated. These consist of two homogeneous fractions PS-I and PS-II. PS-I contains only D-glucose as the monosaccharide constituent. From methylation analysis and periodate oxidation studies, followed by GLC-MS analysis the linkages, the sugar units in PS-I were identified as (1-->3)-D-Glcp and (1-->6)-D-Glcp. PS-II contains D-glucose, and the mode of linkage of d-glucose was identified as (1-->6)-D-Glcp. Finally, the following possible structures of the polysaccharides were assigned using 1H, 2D-COSY, TOCSY, NOESY and 13C NMR spectral analysis: [carbohydrate structure: see text].

Agaricales↗
PubMed2004

The structure of the glucuronoxylomannan produced by culinary-medicinal yellow brain mushroom (Tremella mesenterica Ritz.:Fr., Heterobasidiomycetes) grown as one cell biomass in submerged culture.

The yellow brain mushroom Tremella mesenterica possesses a wide spectrum of medicinal properties, including immunostimulating, protecting against radiation, antidiabetic, anti-inflammatory, hypocholesterolemic, hepatoprotective, and antiallergic effects. A unique feature of T. mesenterica is that most of the above mentioned medicinal properties depend on glucuronoxylomannan (GXM) contained in fruiting bodies or produced in pure culture conditions. We developed a new strain of T. mesenterica CBS 101939, which grows in submerged culture and offers superior yields of one-cell biomass rich in exocellular heteropolysaccharide GXM. The structure of the GXM was analyzed by NMR spectroscopy and chemical methods. The polysaccharide has a defined repeating unit structure, which is O-acetylated at several points: [structure: see text]. These results differ from previously published structure of Tremella extracellular polysaccharides, where mannan backbone was believed to be randomly glycosylated with xylan chains of different length.

Agaricales↗
PubMed2004

Structural investigation of a water-soluble glucan from an edible mushroom, Astraeus hygrometricus.

A water-soluble glucan, Fraction I, was isolated from the aqueous extract of the fruit bodies of the mushroom Astraeus hygrometricus. On the basis of total hydrolysis, methylation analysis, periodate oxidation, and NMR studies ((1)H, (13)C, 2D-COSY, TOCSY, NOESY, and HSQC), the structure of the repeating unit of the glucan is determined as:This glucan shows strong splenocyte activation.

Agaricales↗
PubMed2005

Structural investigation of a polysaccharide (Fr. II) isolated from the aqueous extract of an edible mushroom, Pleurotus sajor-caju.

A water-soluble polysaccharide, (Fr. II) isolated from aqueous extract of an edible mushroom Pleurotus sajor-caju, was found to consist of D-glucose, D-galactose, and D-mannose in a molar proportion of 1:1:1. Compositional analysis, methylation analysis, periodate oxidation study, partial hydrolysis, and NMR experiments ((1)H, (13)C, 2D-COSY, TOCSY, NOESY, HSQC, and HMBC) revealed the presence of the following repeating unit in the polysaccharide: [formula: see text]

Carbohydrate Sequence↗
PubMed2005

Useful applications of DOSY experiments for the study of mushroom polysaccharides.

DOSY analysis has been performed on different mushroom extracts and fractions with the aim to describe a general method for the study of high molecular weight polysaccharides. These NMR experiments can be exploited to monitor the fractionation pathways performed on crude extracts in order to isolate polysaccharides. DOSY can also rapidly verify the purity of the isolated compounds, as well as evaluate their molecular size. In spite of the complexity of DOSY spectra of mixtures, this NMR technique seems to be a valid analytical tool that could be adopted as a routine method for the study of polysaccharides from different sources.

Agaricales↗
PubMed2006

Isolation and structural elucidation of a water-soluble polysaccharide (PS-I) of a wild edible mushroom, Termitomyces striatus.

A heteropolysaccharide (PS-I), isolated from the hot aqueous extract of an edible mushroom, Termitomyces striatus, is composed of d-glucose, d-galactose, d-mannose and l-fucose in a molar ratio 2:1:1:1. Structural investigation of the native as well as the Smith-degraded polysaccharide was carried out using methylation analysis, periodate oxidation studies and 1D and 2D NMR spectroscopy, and the repeating unit of the polysaccharide is established as follows: [carbohydrate structure: see text]

Agaricales↗
PubMed2006

The structure of a polysaccharide from Fraction-II of an edible mushroom, Pleurotus florida.

A water-soluble polysaccharide was isolated from Fraction-II of the aqueous extract of the fruit bodies of the mushroom, Pleurotus florida. Compositional analysis, methylation analysis, periodate oxidation study, Smith degradation, and NMR studies (1H, 13C, DQF-COSY, TOCSY, NOESY, HSQC, and HMBC) revealed the presence of the following repeating unit in the polysaccharide: [structure: see text].

Carbohydrate Sequence↗
PubMed2006

A water-insoluble (1-->3)-beta-D-glucan from the alkaline extract of an edible mushroom Termitomyces eurhizus.

A water-insoluble glucan, TEINS has been isolated from the hot alkaline extract of an edible mushroom Termitomyces eurhizus. The total carbohydrate content of the polysaccharide fraction was found to be 98.4%, and it was found to contain only glucose as the monosaccharide constituent. On the basis of total acid hydrolysis, a methylation experiment, periodate oxidation and (13)C NMR experiment, the repeating unit of the polysaccharide was established as: -->3)-beta-D-Glcp-(1-->.

Agaricales↗