An antifungal protein from the culinary-medicinal beech mushroom, Hypsizygus marmoreus (Peck) Bigel. (Agaricomycetideae)

An antifungal protein (HM-af) was purified from the culinary-medicinal mushroom Hypsizygus marmoreus. The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry of HM-af indicated that its molecular mass was 9.5 kDa. The N-terminal amino acid sequence of HM-af showed homology to ribonuclease H from Clostridium thermocellum. HM-af showed the antifungal activity against Flammulina velutipes.     

Partial purification and characterization of polyphenoloxidase from culinary-medicinal Royal Sun mushroom (the Himematsutake), Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae)

The Royal Sun mushroom, the Himematsutake culinary-medicinal mushroom, Agaricus brasiliensis has several polyphenoloxidase activities in a broad sense. Here we report the partial purification of tyrosinase-type polyphenoloxidase (PPO). PPO is purified from A. brasiliensis without browning using a two-phase partitioning with Triton X-114 and ammonium sulfate fractionation. Partially denaturing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide electrophoresis) staining with L-3,4-dihydroxyphenylalanine was performed and the indicated molecular sizes were approximately 70 kDa and 45 kDa. The purified enzyme is in its latent state and can be activated maximally in the presence of 1.6 mM sodium dodecyl sulfate (SDS). This enzyme catalyzes two distinct reactions, monophenolase and diphenolase activity, and the monophenolase activity showed a lag time typical of polyphenoloxidase. The K(m) value for 4-tert-butylcatechol was quite similar in the presence and absence of SDS, but the apparent V(max) value was increased 2.0-fold by SDS. Mimosine was a typical competitive inhibitor with K(i) values of 138.2 microM and 281.0 microM n the presence and absence of SDS, respectively.     

Antitumor effect of culinary-medicinal oyster mushroom, Pleurotus ostreatus (Jacq.: Fr.) P. Kumm., derived protein fraction on tumor-bearing mice models

Previously, we reported the in vitro anticancer and immunomodulatory effect of a protein fraction designated as Cibacron blue affinity purified protein (CBAEP) obtained from the culinary-medicinal oyster mushroom, Pleurotus ostreatus. In the present study, we investigated the in vivo antitumor potential of CBAEP in different tumor-bearing mice models and studied the detailed mechanism of tumor regression in Dalton lymphoma (DL)-bearing mice. The lethal dose (LD50) of CBAEP was found to be 55 mg/kg body weight and sublethal doses (5 mg/kg and 10 mg/kg body weight) showed a prolonged tumor survival time in DL, Sarcoma-180, and B16F0 melanoma tumor-bearing mice. Further, CBAEP reduced about 35.68 and 51.43% DL cell growth in 5 and 10 mg/kg body weight, respectively. The in vivo CBAEP treatment showed an apoptotic feature as demonstrated in morphological study and sub-G0/G1 population in cell cycle and Western blot of DL cells. CBAEP also activated immunosuppression condition in DL tumor-bearing host. It also stimulated immune cells in the presence of nonspecific immunostunulator (LPS and ConA) ex vivo as well as enhanced Th1 response with production of TNF-α, IFN-γ, and IL-2. Moreover, it activated tumor-associated macrophages and NK cells. The present findings revealed the potent antitumor property of CBAEP, which might help in developing a new anticancer drug.     

Expression of hygromycin B resistance in oyster culinary-medicinal mushroom, Pleurotus ostreatus (Jacq.:Fr.)P. Kumm. (higher Basidiomycetes) using three gene expression systems

Three hygromycin B phosphotransferase (hph) gene expression systems for culinary-medicinal Oyster mushroom, Pleurotus ostreatus, plasmid pSHC, pAN7-1, and pBHt1 were evaluated through PEG/CaCl(2)-mediated protoplast transformation. Plasmid pSHC is a newly constructed hph gene expression system, composed of Escherichia coli hph gene, the P. ostreatus sdi promoter, and the CaMV35S terminator. The vector pAN7-1 was commonly used for integrative transformation in filamentous fungi. Plasmid pBHtl is a T-DNA binary vector, usually introduced into fungi by Agrobacterium-mediated transformation. The results showed that plasmids pSHC, pAN7-1, and pBHt1 were all integrated into the host chromosomes and expressed hygromycin B resistance in P. ostreatus. pAN7-1 had the highest transformation efficiency and hph gene expression level, pSHC the second, and pBHt1 the lowest. Growth rates of the transformants on plates containing hygromycin B were in correspondence with their hph gene expression levels. To our knowledge, this is the first report on integrated transformation of plasmid pAN7-1 and pBHt1 in P. ostreatus.     

Properties and potential applications of the culinary-medicinal cauliflower mushroom, Sparassis crispa Wulf.:Fr. (Aphyllophoromycetideae): a review

Sparassis crispa is a culinary-medicinal mushroom that has recently become popular in Korea, China, Japan, Germany, and the USA. S. crispa is a good source of food and nutraceuticals, or dietary supplements, due to its rich flavor compounds and beta-glucan content. This review is a comprehensive summary of its distribution, growth, management, general constituents, functional ingredients, as well as its current and potential medicinal and other applications.     

蜜环菌菌丝体化学成分的研究——Ⅶ.丙酮提取物中化学成分的分离与鉴定

自人工液体发酵生产的白磨科(Tricholomataceae)蜜环菌[Armiilaria meilea(Vahl.exFr.)Quel.]菌丝体的丙酮提取物中分离得到八个化合物,其中三个为原伊鲁烷型倍半萜醇芳香酸酯类化合物。根据光谱(UV,IR,¹H-NMR,¹³C-NMR,2D-NMR和MS)分析,其中一个为已知物4′-methylmelledonal(Ⅰ),两个为新化合物即密环菌丑素(armillaritin)和蜜环菌寅素(armillarivin),其结构分别为式Ⅱ和Ⅲ。另外五个为已知物赤藓醇(D-erythritol),甘露醇(Dmannitol),杜鹃花酸(azclaic acid)、苔藓酸(orsellinic acid)和甘油醇-α-单油酸酯(glycerin-α-monooleate)。     

蜜环菌菌丝体化学成分的研究——Ⅵ.蜜环菌癸素(armillaripin)的分离与结构

从人工发酵培养的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel.属白蘑科(Tricholometaceae)真菌]菌丝体的石油醚提取物中,经硅胶层析分离得到一新的倍半萜醇芳香酸酯类化合物,命名为蜜环菌癸素(armillaripin)。根据光谱(UV,IR,¹HNMR,¹³CNMR和MS)分析推测其化学结构,并进一步用X-线衍射晶体解析法确证其结构和其相对构型。     

原伊鲁烷型倍半萜醇芳香酸酯的碳谱研究

原伊鲁烷(Protoilludane)型倍半萜芳香酸酯类化合物是近十年来自人工发酵的真菌培养的菌丝中分离得到的一类新型天然产物。作者等自蜜环菌(Armillariamellea)菌丝体中分离得到17个该类型化合物,研究这些新化合物的化学结构,对其全去偶碳谱、偏共振谱或 INEPT 谱、~(13)C-~1HCOSY 谱和远程~(13)~1HCOSY 谱详细地进行分析研究,指出每个化合物各碳的归属,讨论了其碳谱的特征,将其应用于有关化合物的结构测定。     

原伊鲁烷型倍半萜醇芒香酸酯的碳谱研究

原伊鲁烷（Protoilludane）型倍半萜芳香酸酯类化合物是近十年来自人工发酵的真菌培养的菌丝中分离得到的一类新型天然产物。作者等自蜜环菌（Armillaria mellea）菌丝体中分离得到17个该类型化合物，研究这些新化合物的化学结构，对其全去偶碳谱、偏共振谱或INEPT谱、^13C-^1H COSY谱和远程^13C-^1H COSY谱详细地进行分析研究，指出每个化合物各碳的归属，讨论了其碳谱的特征，将其应用于有关化合物的结构测定。     

蜜环菌菌丝体化学成分的研究——Ⅴ.蜜环菌辛素和蜜环菌壬素的分离与鉴定

从人工液体发酵培养的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel.]菌丝体的石油醚提取物中,经硅胶柱层析,低压柱层析和制备性薄层层析分离得到两个新的原伊鲁烷型倍半萜芳香酸酯类化合物。命名为蜜环菌辛素(armillarilin)和蜜环菌壬素(armillarinia),根据光谱(UV,IR,¹HNMR,¹³CNMR和MS)分析,推定它们的化学结构分别为Ⅰ和Ⅱ。     

Chemical and biological properties of indole glucosinolates (glucobrassicins): a review

Glucosinolates are a group of secondary products commonly, but not exclusively, found in plants of the family Cruciferae. They give rise, upon enzymic hydrolysis, to a range of volatile, pungent and physiologically active compounds. Recently, particular attention has been focused upon those that are trytophan-derived--the indole glucosinolates (glucobrassicins). When chemically or enzymically hydrolysed these compounds give rise to a range of involatile indole compounds which have been implicated in the anti-carcinogenic and mixed-function-oxidase stimulatory activities of brassica vegetables. This review details the chemical and physiological properties of indole glucosinolates and their products and suggests possible areas for future research.     

Characterization of blood beta-1,3-glucan and anti-beta-glucan antibody in hemodialysis patients using culinary-medicinal Royal Sun Agaricus, Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae)

Beta-glucan is a major component of fungal cell walls and shows various immunopharmacological activities including antitumor activity. Previously, we detected anti-beta-glucan antibody in human sera. Anti-beta-glucan antibody participates in the immune response to fungal cell wall beta-glucan. Patients on dialysis are at high risk of infection including fungal infections. We examined the plasma beta-glucan level and the titer of anti-beta-glucan antibody in dialysis patients. We measured plasma beta-1,3-glucan concentrations with the limulus G test and anti-beta-glucan antibody titers by ELISA with Candida beta-glucan-coated plates. We also examined the influence of the period of dialysis and the kind of dialysis membrane. The patients were positive for beta-1,3-glucan in their plasma. The anti-beta-glucan antibody titer was lower in the dialysis patients than in healthy volunteers. Long-term dialysis patients showed lower anti-beta-glucan antibody titers than short-term dialysis patients. No significant difference was found between the kinds of dialysis membrane. The titer of anti-beta-glucan antibody as recognition molecule of beta-glucan was low in dialysis patients compared with healthy volunteers. This is likely to be one factor explaining the sensitivity to infection of the dialysis patients. An appropriate application of culinary-medicinal mushroom such as Agaricus brasiliensis has potential for the prevention of fungal infection in dialysis patients.     

Nonvolatile taste components of culinary-medicinal maitake mushroom, Grifola frondosa (Dicks.:Fr.) S.F. Gray

Culinary-medicinal maitake mushroom Grifola frondosa (Dicks.: Fr.) S.F. Gray mycelium was inoculated into wheat, a new product was formed after fungal fermentation, and its nonvolatile taste components were studied. Contents of crude ash, fat, fiber, and protein were higher in fruit bodies and mycelia, whereas carbohydrate contents were higher in the uninoculated and fermented wheat. Contents of total soluble sugars and polyols were 234.06, 111.70, 88.58, and 28.83 mg/g for fermented wheat, fruiting bodies, mycelia, and wheat, respectively. Contents of total free amino acids were in the descending order of mycelia > fermented wheat > fruiting bodies > wheat. Contents of total 5'-nucleotides were in the descending order: fruiting bodies > mycelia > fermented wheat > wheat. Based on the results obtained, fruiting bodies and mycelia possessed highly intense umami taste. Overall, as a result of solid-state fermentation, fermented wheat contained more taste components and a higher equivalent umami concentration than wheat did.     

Preparation of culinary-medicinal maitake mushroom, Grifola frondosa (Dicks.: Fr.) S.F. Gray (Aphyllophoromycetideae)-fermented wheat and its antioxidant properties

The optimization of solid-state fermentation conditions for mycelial growth in wheat by culinary-medicinal maitake mushroom, Grifola frondosa (Dicks.: Fr.) S.F. Gray was studied. The results from the center of the design analysis showed that the recommended optimal conditions for carbon, nitrogen sources, and temperature were 19 mg lactose/g, 2.0 mg malt extract/g, and 30 degrees C, respectively. Under these conditions, the mycelial biomass of 533.1 mg/g was obtained in fermented wheat, comparable to that (545.7 mg/g) obtained using the one-factor-at-a-time method. Based on the results obtained, EC50 values of ethanolic and hot water extracts from fermented wheat were 1.13-15.27 mg/mL, indicating that fermented wheat was effective in antioxidant activity, reducing power, scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals, and chelating ability on ferrous ions. Total phenol contents of both extracts were 13.35 and 23.01 mg/g, respectively. Accordingly, the information obtained would be applicable for the cultivation and consumption of G. frondosa-fermented wheat to provide its beneficial health effects.     

Chemical characterization and biological activity of Macfadyena unguis-cati (Bignoniaceae)

Macfadyena unguis-cati (L.) has been widely used in folk medicine as an anti-inflammatory, antimalarial and antivenereal. The purpose of this study was to chemically characterize the main plant components, and to evaluate the biological properties of some of the fractions derived from leaves (MACb) and liana (MACa) of this plant. Chemical characterization allowed the identification of the compounds corymboside, vicenin-2, quercitrin, chlorogenic acid, isochlorogenic acid, lupeol, beta-sitosterol, beta-sitosterylglucoside, allantoin and lapachol. The biological screening of fractions and/or purified substances derived from fractions revealed antitumoral and antitrypanosomal activities in fractions MACa/lapachol and MACb/MACb21, respectively. The anti-lipoxygenase and anti-cyclooxygenase effect seen in fractions MACa and MACb showed a partial correlation with the anti-inflammatory property attributed to this plant.     

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.     

Optimization of cultivation conditions of fermented shaggy ink cap culinary-medicinal mushroom, Coprinus comatus (O.Mull.:Fr.) Pers. (higher Basidiomycetes) rich in Vanadium

The present paper is mainly aimed at optimization of cultivation conditions of fermented mushrooms of Coprinus comatus rich in vanadium (CCRV). Initial screening of effects of carbon source, temperature, pH, and inoculum size were done by using a one-factor-at-a-time method. The results obtained in that study showed that the optimal medium composition was 30 g glucose/Lin YEPG medium, initial pH 6.0, inoculum volume 10%, and incubation time 120 h. Then the medium was subjected to screening of the most significant parameters using the L9 orthogonal array to solve multivariable equations simultaneously. The results obtained in this study showed that the optimal medium composition was 0.4% V and 30 g glucose/Lin YEPG medium, initial pH 5.0, inoculum volume 15%, and incubation time 120 h. At this medium composition, the mycelial biomass and V content were 7.18 ± 0.24 g/L and 3786.0 ± 17 μg/g, respectively. The anti-diabetic potential of CCRV produced with the optimal level was tested in alloxan-induced diabetes. After the mice were administered (i.g.) with CCRV, the level of blood sugar in the CCRV group was very close to that of the control group. These findings suggested that CCRV produced with the optimal level is useful in the control of diabetes mellitus.     

St. John's wort (Hypericum perforatum): a review of the current pharmacological, toxicological, and clinical literature

RATIONALE: St. John's wort (Hypericum perforatum) has recently gained popularity as an alternative treatment for mild to moderate depression. Given the current widespread use of this herbal remedy, it is important for medical professionals to understand the potential pharmacological pathways through which Hypericum may exert an antidepressant effect. OBJECTIVES: (1) To review the current pharmacological, toxicological, and clinical literature available on Hypericum, and (2) to provide a synthesis of this information into a form that may be easily used by health care providers. METHOD: A comprehensive review of the recent scientific literature (January 1990-March 2000) was performed using the following electronic databases and reference publications: MEDLINE, The Cochrane Library, HealthSTAR, Current Contents (all editions), European Scientific Cooperative on Phytotherapy monographs, German Commission E monographs, and the Physicians' Desk Reference for Herbal Medicines, 1st edition. RESULTS: One hundred and seven (107) publications in the English language and three publications in German were included in the review. Collectively, the data suggest that therapeutic preparations of Hypericum extract appear to exert potentially significant pharmacological activity within several neurochemical systems believed to be implicated in the pathophysiology of depression. However, little information exists regarding the safety of Hypericum, including potential herb-drug interactions. CONCLUSIONS: Additional research on the pharmacological and biochemical activity of Hypericum and its several bioactive constituents is necessary to further elucidate the mode(s) of antidepressant action. Given what is currently known and unknown about the biological properties of Hypericum, those who choose to use this herb should be closely monitored by a physician.     

Hypolipidemic and antiatherogenesis effect of culinary-medicinal pink oyster mushroom, Pleurotus salmoneostramineus L. Vass. (higher Basidiomycetes), in hypercholesterolemic rats

The present study was conducted to investigate dietary supplementation of Pleurotus salmoneostramineus fruiting bodies on biochemical and histological effects in hyper- and normocholesterolemic rats. Six-week-old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Feeding of diet containing a 5% powder of the fruiting bodies of P. salmoneostramineus in hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein, total lipid, phospholipids, and LDL/HDL ratio by 22.55, 51.38, 69.23, 29.67, 16.61, and 65.31%, respectively. The mushroom also significantly reduced body weight in hypercholesterolemic rats. Moreover, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, and enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. salmoneostramineus significantly reduced plasma β and pre-β-lipoprotein, while it increased α-lipoprotein. A histological study of liver tissues by conventional hematoxylin-eosin and oil red O staining showed normal in mushroom feed hypercholesterolemic rat. This study suggests that the P. salmoneostramineus diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.