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The medicinal fungus Phellinus linteus, in the family Hymenochaetaceae, has been used as a traditional medicine for the treatment of various diseases. In this study, the chemical constituents of the culture broth of P. linteus were investigated. P. linteus was cultured in potato dextrose broth medium, and the culture broth was extracted with ethyl acetate. The ethyl acetate-soluble portion was concentrated and subjected to ODS column chromatography, followed by Sephadex LH-20 column chromatography. Six compounds (1~6) were purified by preparative reversed-phase high-performance liquid chromatography. Spectroscopic methods identified their structures as caffeic acid (1), inotilone (2), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (3), phellilane H (4), (2E,4E)-(+)-4'-hydroxy-γ-ionylideneacetic acid (5), and (2E,4E)-γ-ionylideneacetic acid (6). Compounds 1, 2, and 3 exhibited potent dose-dependent antioxidant activity.

Spawn is the vegetative growth of the mycelium and serves as the inoculum or seed for cultivating mushrooms. Various cereal grains, such as wheat, millet, barley, sorghum, brown rice, rye, and oat were assessed to compare their characteristics and suitability for spawn production of button mushroom (Agaricus bisporus). The mycelial growth rates, density, and the number of completely colonized grains were measured from the twentieth day of inoculation. Wheat grains showed fastest mycelial growth with 8.4 cm followed by rye, oat, barley with 8.2, 7.5 and 7.3 cm, respectively. In the mycelial density, foxtail millet, barley, and sorghum were best compared with that of wheat grains. Especially, the number of grains which were completely colonized by mycelia were greatest in foxtail millet with 5,123 grains followed by proso millet, and wheat with 3,052 and 914, respectively. Based on the results obtained, barley, foxtail millet, and sorghum grains would be appropriate substituting for wheat grain in spawn production of button mushroom.

This study was aimed to find factors responsible for export potential of oyster mushroom through postharvest treatments including high pressure and anti-moisture pad. Temperature fluctuation during export simulation induced the condensation of excess moisture resulting in the quality loss due to browning and decay. Anti-moisture pad was effective on inhibition of browning occurrence in part. High temperature exposure () as retail simulation greatly offset the positive effect of anti-moisture pad. Short term treatment of high pressure delayed the surface color changes as hunter L, a, and b values. There was no consistent tendency in total sugar and phenolics between treatments. In general, the export potential of small size mush-room (export size) at harvest was higher than large ones (domestic size). The major factors asso-ciated with the quality decrease of oyster mushroom during export were moisture condensation in packages and temperature fluctuation. Further research is required to improve mushroom export procedures.

Four Cladobotryum isolates were collected from four different commercially grown mushroom types infected with cobweb disease in Cheongdo-gun and Chilgok-gun of Gyeongbuk Province, Korea in 2010. The isolates were identified as C. mycophilum from Agaricus bisporus and Pleurotus eryngii, C. varium from Flammulina velutipes and Hypsizygus marmoreus. The cultural characteristics of the four isolates were investigated using potato dextrose agar (PDA) media under nine different temperatures ranging from 5~32℃. Rapid growth of the isolates to colony diameters of 47~82 mm was observed at conditions of 18~22℃. No growth was observed at 32℃. C. mycophilum produced a yellowish red pigment while C. varium produced a cream colored pigment after cultivation for 25 days on PDA. Phylogenetic analysis of the internal transcribed spacer region and partial 28S rDNA from the four isolates confirmed they were C. mycophilum and C. varium. Cross pathogenicity tests revealed that the two isolates of C. mycophilum were highly pathogenic toward three mushroom types, but not toward H. marmoreus. The two isolates of C. varium were less pathogenic than those of C. mycophilum, but were pathogenic toward all mushroom types evaluated.

In order to apply into functional food or medicinal industry, nutritional characteristics and physiological functionality of Hypsizygus marmoreus (brown cultivar) were investigated. Fruiting body of H. marmoreus contained 27.3% of crude protein, 55.8% of total sugar and 11,109.3 mg/100 g dry weight of malic acid. Furthermore, 66.7% of antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity and 37.3% of antigout xanthin oxidase inhibitory activity showed in the water extract from H. marmoreus. The economically ACE inhibitory activity (81.4%) was obtained when the fruiting body of Hysizygus marmoreus was extracted with distilled water of 50 o C for 12 h, even though maximal ACE inhibitory activity (84.4%) was showed the extracts from 70 o C for 12 h.

This study investigated the inhibitory effects of oligoporin A on platelet aggregation and the mechanism of its action on downstream signaling molecules. Oligoporin A was isolated from the fruiting bodies of Oligoporus tephroleucus (Polyporaceae). The anti-platelet activities of oligoporin A were studied using rat platelets. The effects of oligoporin A on intracellular Ca(2+) mobilization, ATP release, production of the cyclic nucleotides cAMP and cGMP, extracellular signal-regulated kinase (ERK) 2 phosphorylation, and fibrinogen binding to active integrin α(II)(b)β(3) were assessed. Oligoporin A, but not oligoporins B and C, inhibited collagen-induced platelet aggregation in a concentration-dependent manner. Interestingly, oligoporin A did not affect ADP- and thrombin-induced platelet aggregations, which act on different types of membrane receptors. Granule secretion analysis demonstrated that oligoporin A significantly and dose-dependently reduced collagen-induced ATP release and intracellular Ca(2+) mobilization. Additionally, oligoporin A induced the dynamic increase in cAMP and cGMP. Increased cGMP production was further confirmed by the simultaneous production of nitric oxide. Pretreatment with oligoporin A significantly blocked collagen-induced ERK2 phosphorylation. Finally, oligoporin A vaguely diminished the binding of fibrinogen to its cognate receptor, integrin α(II)(b)β(3). The results indicate that oligoporin A inhibits only collagen-induced platelet aggregation mediated through the modulation of downstream signaling molecules. Oligoporin A may be beneficial against cardiovascular disease provoked by aberrant platelet activation.

JK BioTech Co. Ltd. Gyonggi, Ansung 456-749, Korea(Received 16, Novembr 2011., Accepted 21, November 2011)ABSTRACT: This study was performed to develop new Hypsizygus marmoreus cultivars that have enhanced functionalmaterials and improved physiological characteristics with mutagenesis by gamma ray irradiation. Protoplasts of H.marmoreus brown strain HYM-056 were irradiated by gamma ray for mutagenesis, and then 2,000 clones of mutantswere randomly selected and the fruiting bodies were induced by bottle culture. Among them, 157 isolates with fast-growing,heavy and many fruiting body-producing were selected. The isolates were cultured in plastic bottle containing ricebran, barley hulls and fir sawdust to form the fruiting bodies. About 100 days after inoculation, characteristic of fruitingbodies were investigated. The isolates were divided into 6 groups based on color, shape and size of pileus, and length,diameter, number and weight of stipe. In addition, the genetic variation of the isolates was analyzed by URP-PCRfingerprinting.KEYWORDS : Gamma-ray irradiation, Hypsizygus marmoreus, Mutagenesis

To develop health food and alternative medicine, water and ethanol extracts from Hypsizygus marmoreus (white cultivar) fruiting body were prepared, and its physiological functionalities were investigated. Antihypertensive angiotensin I-converting enzyme (ACE) inhibitor activity from water extract was showed higher of 60.5% than ethanol extract and SOD-like activity and xanthine oxidase inhibitory activity were also showed 24.1% and 23.0%, respectively. The other functionalities were very low or not detected. The maximal ACE inhibitory activity (80.5%) was obtained when the fruiting body of Hypsizygus marmoreus was extracted with distilled water (dilution 1 : 30) at for 12 h.

야생 능이버섯에 함유되어 있는 생리 기능성 물질들을 탐색, 분리하여 이들을 고부가가치의 건강식품 소재로 활용하고자 덕유산 일대에서 채집한 능이버섯의 물 추출물은 제조하여 몇 가지 생리 기능성을 측정하였다. 능이버섯 자실체의 물 추출물은 항고혈압성 엔지오텐신 전환효소(ACE) 저해 활성과 항통풍성 xanthine 산화효소(XOD) 저해활성이 각각 74.3%와 59.6%로 우수하였다. 그러나 tyrosinase 저해 활성과 항산화 활성, SOD 유사활성 등은 매우 낮거나 없었다. 능이버섯 자실체중의 ACE 저해활성은 자실체 분말의 물현탁액을 $30^{\circ}C$ 에서 24시간, XOD 저해물질은 $50^{\circ}C$ 에서 24시간 진탕 시켰을 때 가장 높았다. 이들 두 가지 생리활성 물질을 함유한 능이버섯 물 추출물은 앞으로 건강식품이나 대체 의약 소재 개발에 매우 유용하게 활용 할 것으로 사료된다. 【We collected wild mushroom, Sarcodon aspratus from Deogyu Mt. of Muju, Jeollabuk-do and physiological functionalities of its water extract were investigated. The water extracts from S. aspratus showed the highest antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity of 74.3% and also showed high anti-gout xanthine oxidase (XOD) inhibitory activity (59.6%). However, tyrosinase inhibitory activity was very low (17.3%), and antioxidant activity and SOD-likely activity were not detected. The ACE inhibitory activity of S. aspratus fruiting body was the highest when powder of the fruiting body was shaked at $30^{\circ}C$ for 24 h by distilled water. Furthermore, the XOD inhibitory activity was also the highest by extraction at $50^{\circ}C$ for 24 h with water.】

【The purpose of the present study is to develop a simple, fast and sensitive LC/MS method for simultaneous separation and the determination of an active component in the oriental medicinal mushroom Cordyceps militaris. Based on this work, the contents of cordycepin in Cordyceps militaris fruiting cultivated on various media were determined and compared. And also, the nutritional components such as minerals and vitamins were determined in order to provide useful information to consumer as a food material. The analysis methods of nutritional components were chosen on the basis of AOAC. The optimum separation for cordycepin was achieved using a solvent gradient consisting of the mixture of 0.1% formic acid in methanol (solvent B) in a background of 0.1% formic acid in water (solvent A) as a mobile phase and a 3.0 ${\times}$ 150 Waters XTera column. Selective ion monitoring (SIR) mode ([M+H]+ at m/z 252) was used for quantitative analysis of cordycepin. The cultivated Cordyceps militaris on various media contained 1~14 /g of cordycepin, 0.65~1.08% of thiamine, 0.86~7.17% of riboflavin, and 3.01~5.26% of niacin. The content of mineral components varied on categories, especially contained 500~3500% of potassium as a major mineral. Cordycepin, niacin and potassium were found much higher in the fruiting cultivated with soy power media (gold 10) than other media.】

Novel styrylpyrones, phellinins A1 and A2, were isolated together with known styrylpyrone compounds, hispidin and 1,1-distyrylpyrylethan, from the cultured broth of Phellinus sp. KACC93057P. These compounds were purified by solvent partition, Sephadex LH-20 column chromatography, C(18)-solid phase extraction and finally by reversed-phase (ODS) TLC. To identify the phellinin producer Phellinus sp. KACC93057P, the ribosomal DNA (rDNA) internal transcribed space regions containing 5.8 rDNA were sequenced and compared with those of the known Phellinus isolates. Phellinus sp. KACC93057P was 94.8% identical to P. baumii and P. linteus, all of which did not produce phellinins A1 and A2. These compounds significantly scavenged free radicals such as 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) and superoxide.

Effects of concentration (500, 800, 1,000 and 1,500 ppm) on the initiation and development of fruit body of Grifola frondosa on sawdust cultivation were studied. Optimum concentrations of carbon dioxide for the initiation and development of the fruit body showed the ranges from 500 to 800 ppm. Fruit body initiation was accelerated at lower than 800 ppm exposure but the maturing of the fruit body was not influenced by above treated concentrations. The higher ratio of primordial formation, faster fruit body initiation and higher yield were obtained at below 800 ppm of level, whereas over 1,000 ppm of levels showed abnormal and lower quality of fruiting bodies. Based on the above results, it is concluded that the favorable level for bag culture of G. frondosa was below 800 ppm.

This study was conducted to investigate genetic characteristics of 25 Pleurotus eringii strains that have been released in Korea based on cultural, morphological features and PCR fingerprints. Strains PER-007 and PER-012 showed distinct cultural characteristics in growth rate, morphological characteristics of mycelial colony and fruiting bodies when compared to those of other strains. Strain PER-007 did not form primordium initiation in sawdust medium and PER-012 also showed different phenotypes on fruiting bodies. Eleven URP primers were used to detect PCR polymophic bands in P. eringii strains. Primers URP1F, URP2R, URP2F, URP4R, URP6R, URP9F and URP17R were selected as useful primers for amplifying PCR polymorphic bands in P. eringii strains. The genetic similarity index was calculated by using PCR polymorphic bands amplified by eight URP primers among the 25 strains. The P. eringii strains were grouped by four distinct clusters on the UPGMA analysis. The genetic similarity values ranged from 100% to 76% were observed in three major groups, suggesting close genetic relatedness of them. Exceptionally, PER-007 and PER-017 were involved in outgroup.

This study was conducted to investigate the changes in characteristics of the Phellinus gilvus mushroom as influenced by drying methods after harvest. The lowest weight loss rate of P. gilvus mushroom was 75.8% with drying in the shade and 80% by dryer (60°C). The size loss rate of pileus was 19.3% of that in a hot air dryer (60°C). The hardness of dried material context using a hot air dryer (60°C) was the lowest (20 kg/cm2), and that by a dry oven (60°C) was the highest (457 kg/m2). For ΔE value, 4.9 of context and 2.6 of tubes using drying in the shade (20°C) were found to be the lowest. The survival rate of sarcoma 180 treated with P. gilvus dried in the sun was the lowest (51.8%), and this was considered the most effective method for antitumor activity against sarcoma 180.

To elucidate optimum light conditions for artificial cultivation of Grifola frondosa, the effects of light quality (blue, green, white) and light intensity (200, 500, 800, 1200 lux) on primordium formation, morphological properties and yield of fruiting bodies of G. frondosa using bag cultivation method were tested. Among three light sources, white light source () had a higher mushroom yield (242 g/bag) and a shorter cultivation period (52 days) than those of the others. In particular, blue light source () induced the morphology of wide and deep colored pilei in G. frondosa fruiting body. The experimental results on the appropriate light intensity indicated that 500 lux light was the most effective on mushroom production, whereas primordium formation was effective at 200 lux.

This study was carried out to develop artificial culture method of Grifola frondosa in polypropylene bag. To find out the favorable substrate formulation of G. frondosa, physicochemical conditions were investigated. The T2 formulation (55 : 25 : 12 : 8) mixing ratio of oak sawdust, oak chip, dried bean-curd refuse and wheat bran showed the shortest time to complete the crop cycle and the highest yield (weight of fresh mushrooms harvested at maturity). Those physicochemical conditions were pH 4.3, 2.4% crude fat contents, 54.1% total carbon, 1.42% total nitrogen, 38.1 C/N ratio, 75.5% porosity and bulk density. Among the physicochemical factors, pH, crude fat and total nitrogen may affect mushroom yield. Therefore, development of favorable substrate would be benefit to increase production efficiency of G. frondosa, mushrooms and be commercial potential.

Twelve URP primers were used to assess genetic characteristics of oyster mushroom including 59 Pleurotsu ostreatus cultivars, two of P. florida cultivars, one P. sajor-caju cultivars, one P. abalonus cultivar and two P. eryngii cultivars registered in Korea. Six URP primers produced PCR polymorphic bands within and between the Pleurotus species. Primer URP2F produced distinct cultivar specific PCR polymorphic bands that profiled to 15 cultivar types. PCR polymorphic bands amplified by URP2F, URP6R, URP4R and URP2R were used for UPGMA cluster analysis. Fifty nine cultivars of Pleurotus ostreatus are genetically clustered into 5 groups, showing genetic similarity over 70% among them and P. abalonus. P. eryngii and P. sajor-caju, were involved in outside groups.

This study was carried out to obtain basic data on mycelial growth characteristics for an artificial cultivation of Grifola frondosa. Ten strains of G. frondosa were collected from Korea, China and Japan and investigated its optimal culture condition. Among four kinds of mushroom culture media, PDA medium was selected as the suitable culture medium. The optimal conditions for the mycelial growth of G. frondosa in PDA medium were and of pH, respectively. The carbon and nitrogen sources for the optimum mycelial growth were fructose and peptone, respectively, and the highest mycelial growth was observed when the C/N ratio was .

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.

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 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 for 12 h by methanol and its HMG-CoA reductase inhibitory activity was 80.2%.

Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

A total of 179 isolates of Trichoderma spp. were collected from oyster mushroom substrates in Korea. On the basis of morphological and cultural characteristics, Trichoderma isolates were divided into seven groups, namely T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, referred to as Trichoderma sp. 1 and 2. The predominant species was Trichoderma sp. 2 (n

느타리버섯 균상에 발생하는 Hypocrea속 균의 분류학적 위치를 확립하기 위하여 유성세대 및 무성세대의 형태적 특징을 조사하였고, 국내 분리 균주의 RAPD, RFLP, URP-PCR 분석 등 분자생물학적 특징을 조사하였다. 1. 충남과 전남 일대의 8개 시, 군의 느타리 재배사에서 19 균주를 분리하였으며 이들 균주간의 형태적, 발생생태학적 변이는 없었고, 기존 보고된 Hypocrea 종들과는 다른 형태적 특징을 나타내었다. 2. Hypocrea 자좌의 크기는 $6.0{\sim}13.0{\times}3.0{\sim}11.0mm$ 이었고, 위유조직으로 구성되어 있으며 대부분 군생하고 처음엔 백색이지만 성숙되면 노란빛을 띤 갈색을 나타내었다. 자낭각은 구형으로 크기가 평균 $239.9{\times}204.1{\mu}m$ 이었으며, 자낭에는 16개의 자낭포자(part ascospores)를 형성하였고, 자낭과 장낭포자의 크기는 각각 $103.3{\times}4.9{\mu}m$ 와 $6.5{\times}4.7{\mu}m$ 이었다. 3. 무성세대의 phialide는 보통 Trichoderma virens와 유사한 형태로 모여서 형성되나 때때로 penicillate의 형태도 관찰되었다. T. virens형 phialide의 크기는 $10.8{\times}3.5{\mu}m$ 이었고 penicilliale형은 $14.7{\times}3.4{\mu}m$ 이었다. 분생포자는 계란형에서 타원형으로 표면은 매끈하고 크기는 $4.5{\times}2.9{\mu}m$ 이었다. 4. Hypocrea 균주들의 배양적 특징 조사한 결과, PDA 배지에서 균사생장 최적 온도는 $25^{\circ}C$ 이었고 자좌는 $15{\sim}25^{\circ}C$ 범위에서 발생하였으며 자좌 발생 최적 온도는 $20^{\circ}C$ 였다. 5. RAPD 및 RFLP 분석을 실시한 결과 Hypocrea속 균은 공시균주는 모두 단일 유전 개체군으로 느타리 재배사에서 발생하는 Trichoderma속 균 및 기존에 보고된 Hypocrea spp.와는 다른 분류군으로 나타났다. 【Recently a serious outbreak of weed mould caused by a species of Hypocrea occurred in oyster mushroom (Pleurotus ostreatus) substrates in Korea. The disease was characterized by a rapid infestation of the oyster mushroom substrates by Hypocrea sp. and subsequent inhibition of fructification of the mushroom. In spite of it's serious losses to the oyster mushroom industry in Korea, etiology and ecology of the disease have not been studied. Morphological characteristics of the fungus were examined and molecular characteristics of the fungus were compared with those of the green moulds (Trichoderma spp.) isolated from oyster mushroom bed. Stromata formed superficially on suface of the substrates were pulvinate to effuse or irreguler, initially white but becoming yellowish brown, measuring $6.0{\sim}13.0{\times}3.0{\sim}11.0mm$ . Perithecia were globose to subglobose, immersed in stroma, $223{\sim}263\;(Ave.239.9){\times}167.3{\sim}231\;(Ave.204.1){\mu}m$ in size. Asci were unitunicate, cylindrical, nonamyloid, $82.7{\sim}124.8\;(Ave.103.3){\times}4.1{\sim}5.1\;(Ave.4.9){\mu}m$ in size, 16 part-spored. Ascospores were bullet-shaped or somewhat oblong, hyaline, bicellular, roughened or warted, $5.4{\sim}7.4\;(Ave.6.5){\times}3.6{\sim}5.5\;(Ave.4.7){\mu}m$ in size. This fungus readily form the stroma on PDA. Mycelia on PDA nearly invisible and without cottony aerial mycelium. Optimum temperature for mycelial growth of this fungus was $25^{\circ}C$ on PDA and its growth rate was 15 mm per day. This species did not grow at below 10 and above $35^{\circ}C$ . Phialides in culture enlarged in the middle and aggregated to penicillate type. They were very variable, shorted ampulliform and occasionally curved when matured, but cylinderical when young, measuring $11.9{\sim}24.3\;(Ave.\;14.7){\times}2.9{\sim}3.9\;(Ave.\;3.4){\mu}m$ when matured and $7.2{\sim}14.0\;(Ave.\;10.8){\times}2.8{\sim}4.9\;(Ave.\;3.5){\mu}m$ when young. Phialosopres were ovoid to ellipsoid, smooth, measuring $3.5{\sim}7.2\;(Ave.\;4.5){\times}2.6{\sim}3.3\;(Ave.\;2.9){\mu}m$ . Nineteen isolates of Hypocrea sp. were analyzed on the basis of molecular characteristics and classified into phenotypic groups. On the basis of RAPD, URP-PCR, the fungus was confirm to monoclonal, and was classified as a different taxon from reported species of Hypocrea and Trichoderma and supposed to be a new species not previously reported in literature.】

During a search for neuraminidase inhibitors derived from medicinal fungi, we found that the fermentation broth of Phellinus linteus exhibited potent neuraminidase inhibitory activity. Through bioassay-guided fractionation, two active compounds were purified from the ethyl acetate-soluble portion of the fermentation broth of P. linteus. These structures were identified as inotilone (1) and 4-(3,4-dihydroxyphenyl)-3-buten-2-one (2) by spectroscopic methods. Compounds 1 and 2 inhibited H1N1 neuraminidase activity with IC50 values of 29.1 and 125.6 µM, respectively, in a dose-dependent manner. They also exhibited an antiviral effect in a viral cytopathic effect reduction assay using MDCK cells. These results suggest that compounds 1 and 2 from the culture broth of P. linteus would be good candidates for the prevention and therapeutic strategies towards viral infections.

This study was performed to develop anti-obesity Phellineus linteus powder for application as food additives. P. linteus powders which contained anti-obesity lipase inhibitor were prepared by freeze drying and spray drying. Effects of various additives on the quality of their powders were investigated. 10% over addition of dextrin and gum-arabia was good in the rheological properties of their powders and optimal spray temperature was near . When their powders were sealed and storaged at room temperature for 6 months, its characteristics were not changed. Especially, changes of its colors was repressed by addition of 0.3 M oxalic acid, citric acid and ascorbic acid than those by non-treatment.

Anti-gout xanthine oxidase inhibitory activity of water extracts from various mushrooms were determined. The highest xanthine oxidase inhibitory activity was 72.9% in the water extract from fruiting body of Agaricus brazillensis and also were high in the extract from fruiting bodies of Pleurotus salmoneostramineus(60.1%), Phellinus baumii(57.7%), Agaricus bisporus(56.7%) and Hericium erinaceum(53.4%). The xanthine oxidase inhibitor was maximally extracted when Agaricus brazillensis fruiting body was treated with water at for 24 h.

To elucidate the composition of phytosterols in the fruit body of Pholiota spp. 6 species(P. adiposa, P. aurivella, P. highlandensis, P. nameko, P. squarrosa and Pholiota sp.) were analyzed with Gas chromatography(GC). Pholiota spp. were contained campesterol, stigmasterol, and -sitosterol as the major phytosterols.

Hypertension is one of the very serious diseases and, recently, hypertensive patient longevity has been increased significantly. Therefore, the development of new antihypertensive drugs or bioactive compounds is very important to remedy or prevent hypertension. The antihypertensive angiotensin I-converting enzyme (ACE) inhibitor in water extracts from the brown-cultivar-fruiting-body ofHypsizygus marmoreuswas purified with ultrafiltration, C18solid phase extraction chromatography and reverse-phase HPLC, and the purified ACE inhibitor with inhibitory activity of IC50value of 0.19 mg/mL was obtained. The purified ACE inhibitor was found to be a new oligopeptide with the sequence LSMGSASLSP. Its molecular weight was estimated to be 567.3 Da and the water extracts containing ACE inhibitor fromHypsizygus marmoreusshowed a clear antihypertensive action a spontaneously hypertensive rat.

Ganoderma lucidum develops atypical fruiting structures (AFSs) with non-basidiocarpous basidiospores during the incubation under light on nutrient agar media. To examine the light quality effective in inducing AFSs, 17 isolates ofG. lucidum were incubated on agar media under light from different colored fluorescent lamps. Of the 17 isolates, 13 isolates produced AFSs and basidiospores under fluorescent lamps. Nine isolates formed AFSs in a broad light region from P-B (pure blue) to P-R (pure red) lamps. The remaining 4 isolates produced AFSs under different colored fluorescent lamps. No isolates formed AFSs in the dark or under BLB (black light blue) illumination. The mycelial growth was inhibited by light illumination, especially BLB light. Although the AFSs were induced at a very low light intensity such as 0.5µmol m−2s−1, the optimum light intensity for the AFS formation varied depending on the kind of fluorescent lamp and the isolate. The AFS formation inG. lucidum isolates was also tested under monochromatic light produced by the combination of interference filters and colored glass filters.G. lucidum isolates were separable into various types in the responses of AFS formation to monochromatic light, indicating thatG. lucidum is heterogeneous in its photo-response with regard to AFS formation.

Effects of light and ventilation on the formation of atypical fruiting structures (AFSs) and fruit body primordia (FBPs) ofGanoderma lucidum on nutrient agar media were investigated. Although the mycelial growth was inhibited by illumination and ventilation, brown AFSs appeared on the white mycelial colony, and basidia and basidiospores were produced on the AFSs. On the other hand, FBPs were induced by illumination alone, regardless of ventilation. However, the primordia could not develop to mature fruit bodies. In the dark, only vegetative growth of the fungus progressed. Twenty-three isolates ofG. lucidum collected from four countries were tested for the formation of AFSs and FBPs under light and ventilation. Thirteen isolates formed AFSs, and another five isolates produced FBPs. Of the remaining five isolates, one formed callus-like structures without elaborating basidiospores, and the other four did not induce AFSs or FBPs. Microscopical observation showed that the basidia were formed directly from generative hyphae on the surface of AFSs. Basidiospores formed on the basidia were brown and ellipsoid with an eccentric hilar appendix on the rounded spore base. They had a double wall and mostly contained one or two large vacuoles. The surface of basidiospores was smooth or wrinkled and had shallow holes. The spore size was (4.5−)6.4−9.6(−10.3) × (2.6−)3.2−5.1(−6.4), 7.3 × 4.2 µm on average.

Ginsenoside Rg3, a single ginseng saponin, is known to be a major anti-platelet component of protopanaxadiol that is isolated from Korean red ginseng. In this study, we investigated whether dihydroginsenoside Rg3, a stable chemical derivative of ginsenoside Rg3, also demonstrated anti-platelet activity. Dihydroginsenoside Rg3 inhibited thrombin-induced platelet aggregation in a concentration-dependent manner with an IC50 (concentration producing 50% inhibition) of 18.8 ± 0.4 μM. Ginsenoside Rg3 inhibited platelet aggregation which was induced by thrombin (0.1 U mL−1) with an IC50 of 40.2 ± 0.9 μM. We next determined whether dihydroginsenoside Rg3 affected different types of ligand-induced platelet aggregation. We found that dihydroginsenoside Rg3 inhibited collagen-induced platelet aggregation with an IC50 of 20.0 ± 0.9 μM. To elucidate the inhibitory mechanism of dihydroginsenoside Rg3 on aggregation, we analysed its downstream signalling pathway. It was interesting to note that dihydroginsenoside Rg3 elevated cyclic AMP production in resting platelets, but did not affect cyclic GMP production. In addition, we found that dihydroginsenoside Rg3 potently suppressed phosphorylation of extracellular signal-regulated kinase 2 (ERK2), which was stimulated by collagen (2.5 μg mL−1), but not of p38 mitogen-activated protein kinase. Taken together, our results indicate that dihydroginsenoside Rg3 potently inhibited platelet aggregation via the modulation of downstream signalling components such as cAMP and ERK2.

Dementia is thought to result from β-secretase-mediated precipitation of β-amyloid in the brain. Senile plaque β-amyloid is termed β-amyloid precursor protein (βAPP) (Haass and Selkoe, 1993). The amyloid precursor protein (APP) is processed into many different forms through a combination of cellular protease activities (α-, β-, and γ-secretase). β-Secretase is an aspartic protease also known as BACE1 (the β-site APP-cleaving enzyme). Its activity is the rate-limiting step in Aβ peptide production in vivo (Vassar et al., 1999). β-Secretase has maximal activity at an acidic pH, as agents that disrupt intracellular pH also inhibit β-secretase activity (Knops et al., 1995). β-Secretase is insensitive to pepstatin, an inhibitor of many types of aspartic protease (Sinha et al., 1999). Many groups have attempted to identify β-secretase inhibitors using high-throughput screening of natural product extracts. Peptidic β-secretase (aspartic protease memapsin-2) inhibitor, OM 99-1, and other aspartic protease inhibitors such as OM99-2 and OM99-3 were developed from these efforts (Dorrel, 2000). In addition, non-peptidomimetic derivatives have been synthesized to optimize BACE1 inhibition (Garino et al., 2006). Recently, small-sized synthetic inhibitors containing a tetrazole ring and acidic heterocycle bioisosteres such as KMI-570, KMI-684, KMI-420, and KMI-429 were synthesized (Kimura et al., 2006). In contrast, efforts to discover naturally occurring BACE1 inhibitors have been comparatively limited. Several hydroxyl-containing inhibitors have been reported (Stachel et al., 2004). Catechins from green tea (Jeon et al., 2003), ellagic acid and punicalagin from pomegranate (Kwak et al., 2005), hispidin from mycelial cultures of Phellinus linteus (Park et al., 2004), octapeptide from Saccharomyces cerevisiae (Lee et al., 2007), and several compounds isolated from Sanguisorbae radix (Lee et al., 2005) have all been studied as BACE1 inhibitors. Although the treatment of Alzheimer's disease has been intensively studied, efficacious antidementia drugs or nutraceuticals lacking side effects have yet to be developed. Mushrooms are nutraceuticals with health-stimulating properties and medical effects (Jeong et al., 2004; Kweon et al., 2002; Lee et al., 2003; Park et al., 2003; Yang et al., 2004). Lentinula edodes can be found in Korea, Japan, and China. Few studies have been done regarding the pharmaceutical effects of L. edodes, except for those concerning the cholesterol-lowering effect of eritademine (Kweon et al., 2002), antitumor activity of lentinan (Chung, 1982), and fibrinolytic effect of lentinacin (Chibada et al., 1969). Some commercial blood circulation enhancers as well as a number of antibiotics and immune-stimulating compounds that contain L. edodes have also been developed. The present study was performed to screen varieties of mushroom for a potent antidementia β-secretase inhibitor and to optimize the extraction protocol of the compound, with the aim of developing a novel antidementia β-secretase inhibitor that can be utilized as a drug or in functional foods. The mushrooms used in this study were obtained from the National Institutes of Agriculture Science and Technology (Suwon, Korea) and the Korea National Agricultural College (Hwasung, Korea). Unless otherwise specified, all chemicals and solvents were of analytical grade. The recombinant human BACE1 assay kit was purchased from PanVera (Madison, WI, USA). Dried fruiting bodies (5 g) of mushrooms were pulverized and extracted with 200 ml each of water and methanol at 30℃ for 12 h. The extracts were centrifuged at 10,000 ×g for 20 min and filtered through Whatman No. 41 filter paper. Each supernatant was lyophilized for analysis. The BACE1 inhibitory activity assay was carried out according to the manufacturers protocol with previously described modifications (Byun et al., 2005; Kwak et al., 2005; Vassar et al., 1999). A mixture of 10 µl assay buffer (50 mM sodium acetate, pH 4.5), 10 µl BACE1 (1.0 U/ml), 10 µl substrate (750 nM Rh-EVNLDAEFK-Quencher in 50 mM ammonium bicarbonate), and 10 µl of sample dissolved in the assay buffer was incubated for 60 min at 25℃ in darkness. The mixture was illuminated at an excitation wavelength of 530 nm and light emitted at 590 nm was collected. The percent inhibition was determined by the following equation: [1 - {(S - S0)/(C - C0)}] × 100 where C denotes the fluorescence of a control (enzyme, assay buffer, and substrate) after 60 min of incubation, C0 is the fluorescence of the control at zero time, S is the fluorescence of the tested samples (enzyme, sample solution, and substrate) after 60 min of incubation, and S0 is the fluorescence of the tested samples at zero time. To check the quenching effect of the samples, the sample solution was added to the reaction mixture C, and any reduction in fluorescence by the sample was determined. The IC50 value was defined as the concentration of the BACE1 inhibitor required to reduce 50% of BACE1 activity. All data represent mean values of triplicate experiments. To select the most potent BACE1 inhibitor-containing mushroom, various extracts from 80 species of mushroom fruiting bodies were tested for their BACE1 inhibitory activities (Table 1). Generally, methanol extracts showed higher BACE1 inhibitory activities compared to water extracts. Among the mushrooms tested, methanol extracts of the Lentinula edodes fruiting body showed the highest BACE1 inhibitory activity of 40.1%, and so was selected for further study. L. edodes has long been consumed because of its good rheological properties and nutrition. It also produces fibrinolyltic agents (Chibada et al., 1969) and anticancer compounds (Chung, 1982). The present paper is the first to demonstrate that L. edodes produces a potent antidementia BACE1 inhibitor that may be useful medicinally and as a food supplement. Table 1 β-Secretase inhibitory activities of various extracts from fruiting body of the second-selected mushrooms (unit:%) The effects of temperature on the extraction of the BACE1 inhibitor was investigated. Maximal extraction of the inhibitor was obtained at 40℃, with a BACE1 inhibitory activity of 39.5% evident at this temperature. Additionally, the BACE1 inhibitor was also extracted amply at 50℃ and 70℃, with β-secretase inhibitory activities of 29.0% and 23.0%, respectively, evident. However, at temperatures below 30℃ extraction was below 20% (data not shown). The optimum extraction temperature in the methanol-based procedure is lower than that methanol-based extraction of a platelet aggregation inhibitor from Inonotus obliquus (80℃) (Hyun et al., 2006) but higher than that of Pholiota adiposa HMG-CoA reductase inhibitor (30℃) (Yu et al., 2007). The time course of the BACE1 inhibitor extraction was determined using 50% and 80% methanol. Inhibitor extraction increased as the extraction time increased. The maximal BACE1 inhibitor extraction levels was obtained by extraction after 24 h (Fig. 1). Fig. 1 Effect of extraction time on extraction of BACE1 inhibitor form Lentinula edodes by methanol at 40℃. ▪: 80% methanol extraction, ♦ 50% methanol extraction. The present results show that the optimal extraction condition for extraction of L. edodes BACE1 inhibitor is 50% methanol at 40℃ with an extraction time of 24 h. Under these conditions BACE1 inhibitory activity is 48.5%.

Present experiments were conducted to determine the possibility of artificial culture with various sawdust of P. gilvus. The pH value was 6.0 of oak sawdust, 6.5 of mulberry sawdust, 6.6 of elm sawdust, 6.3 of acacia sawdust and 6.1 of apple tree sawdust. Mycelial density on elm sawdust and acacia sawdust were lower than those of oak sawdust, and apple sawdust. Weight of fresh fruiting body showed that 179 g on oak tree, 227 g on oak sawdust, 21 g on elm tree, 76 g on elm sawdust, 106 g on apple tree, and 170 g on apple sawdust. Among them, the yield of oak substrates was the highest whereas acacia sawdust was the lowest, and it is concluded that the yields of sawdust substrates were higher than log substrates. P. gilvus grown on various sawdusts and logs used in this study have shown similar in anti-tumor activity against P388.

Phellinus genus belonged to Hymenochaetaceae of Basidiomycetes and has been well known as one of the most popular medicinal mushrooms due to high antitumor activity. This study was carried out to obtain the basic information for mycelial culture conditions of Phellinus linteus, P. baumii, and P. gilvus. According to colony diameter and mycelial density, the media for suitable mycelial growth of them were shown in MEA, glucose peptone, and MCM. The optimum temperature for mycelial growth was 30℃. Carbon and nitrogen sources were mannose and malt extract, respectively. The optimum C/N ratio was 10 : 1 to 5 : 1 with 2% glucose concentration, vitamin was thiamine-HCl, organic acid was succinic acid, and mineral salt was MgSO4·7H2O.

A Monokaryotic strain G8M without clamp connections was isolated from germinated basidiospore that was obtained from cultivated fruit body. Strain G8M was used as a tester isolate for ‘dikary on-mo...

This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by P...