Earticle

Recently, farmers of mushroom farms in Korea express the difficulty of shiitake cultivation due to the damage of oak log beds by unknown reason. To elucidate the cause of the damage, we investigated and isolated fungi from inside wood chip of the damaged shiitake log bed. Four of green fungi were obtained and their morphology and molecular properties examined. Trichoderma atroviride, Trichoderma citrinoviride, Hypocrea lixii (Trichoderma harzianum), and Hypocrea lutea (Gliocladiumviride) were identified. Hypocarea lutea is newly described in Korea. These species were able to produce extracellular enzymes such as cellulase, pectinase, and xylanse that might degrade wood components of oak logs. This study provides strong evidence that Trichoderma fungi are involved in the recent damages of shiitake log beds In Korea.

From the used sawdust medium of commercial mushroom, Lyophyllum ulmarium and Pleurotus eryngii, cellulase was extracted by distilled water. Optimum temperature of extraction was 25 ℃. Cellulase including this medium could attack cellulose powder, wood tip of a Japan ceder and cypress, and rice hull. Both cellulase from Lyophyllum ulmarium and Pleurotus eryngii showed high activities for rice hull. Now, by affinity chromatography, these cellulase is tried to purify.

Several bacteria have been known as the causal agents of certain diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). It is well known as bacterial diseases of the cultivated mushroom such as brown blotch, mummy disease, bacterial pit, bacterial rot and weeping disease, ginger blotch, and drippy gill. Brown blotch is the most critical cause of crop loss in the commercial mushroom industry. The classical bacterial blotch disease of mushrooms is caused by a fluorescent pseudomonad, Pseudomonas tolaasii. Affected mushrooms show lesions which become dark chocolate-brown, are wet, and deeply pit the caps and stalks. Although Pseudomonas tolaasii has been known as the casual agent of bacterial blotch, much controversy exists regarding the identification of this bacterium and whether blotch may be caused by more than one organism. This study was carried out to investigate characterization and biological control of Pseudomonas tolaasi and other possible browning pathogens isolated from cultivated mushrooms. One hundred seventy four bacteria were isolated from the cultivated mushroom and collected from main producing districts throughout the country. The isolates were classified into Pseudomonas tolaasii(20 strains), Pseudomonas gingeri(1 strains), Pseudomonas agarici(4 strains), Pseudomonas putida(11 strains), Pseudomonas sp.(46 strains), Ewingella americana(14 strains), Stenotrophomonas sp.(4 strains), and others(74 strains) on the basis of 16 rDNA analysis. The most dominants of these species were Pseudomonas tolaasii and Ewingella americana. Pseudomonad isolates were mainly divided into two groups in white line test and a sharply defined white line of precipitate forms in Pseudomonas agar F(Difco) between the opaque white colonies of P. tolaasii and translucent colonies of certain unidentified pseudomonads. The white line test was positive when 20 isolates of P. tolaasi from different countries were examined, whereas 62 isolates of pseudomonads did not give the white line reaction with a reacting translucent colony Pseudomonas. All the isolates tested for white line forming bacteria including P. tolaasi were highly pathogenic to mushroom tissue. Although browning of mushrooms in host tests does not perfectly help in the identification of P. tolaasi, a conspicuous pitting produced at the cut surface of mushroom tissue is as specific as the white line test in detecting P. tolaasii in suspension in distilled water. URP2F primers of 20-mer were used to assess the genetic diversity of white line forming bacteria. The phylogenetic tree was constructed by using the neighbor-joining method. In the analysis of RAPD pattern, all isolates of white line precipitate have some of the different genetic traits as collected districts. Phylogenetic analysis of 16S rDNA revealed that twenty isolates including white line forming bacteria were closely related to P. tolaasii and showed high similarity. To biological control on bacterial browning disease of cultivated mushrooms, six hundreds plant extracts (332 EtOH extracts, 268 water extracts) was used for control of mushroom disease. Thirty plant extracts in bacterial disease(Pseudomonas tolaasii, P. agarici, B. gladioli, E. americana) and thirty three in fungus disease(T. harzianum, C. mycophilum, V. fungicola) showed strong anti-microbes activity. They showed stronger anti-microbes activity at ethanol extracts than water extracts. MIC of extract BCW128 on Pseudomonas tolaasii was 700ppm and HDE17 was 330ppm. MIC of extract YCE107 on P. agarici was 330ppm, JGE96 was 330ppm and BCW128 was 700ppm. The bacteria inhibit tolaasin secreted by Pseudomonas tolaasii was selected three genus(Bacillus sp. etc). Now we are carrying out more research on these bacteria.

Mushroom is cultivated as one of the major economical crops in many areas of the Korea. The total production have steadily increased approximately 151,913 M/T in 2000 to 186,400 M/T in 2007. This study was carried out to investigate applicability of mushroom production using various organic media resources within the country. Eight organic resources were collected from various areas. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at the media of 10% red ginseng marc, 20% lacquer tree, 20% Juglans mandshurica, 10% Cudrania tricuspidata, 10~20% Acer pensylvanicum, 10% Lindera glauca. Mushroom mycelial growth at red ginseng marc media was slower than that of the control. But the sponin of the red ginseng was not detected at the fruiting body grown from red ginseng marc media, And three organic resources(barly powder, sweet potato powder, potato powder) was used to substitute rice bran used in mushroom cultivation. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at 10~30% sweet potato powder, 20% barly powder and 10% potato powder.

Agaricus bisporus grows on a substrate known as compost, which is a product of aerobic fermentation by various microorganisms. These organisms convert and degrade the straw and form lignin humus complex which is utilized later on by the population of organisms. Theses microflora play a key role in the process of composting and can be regarded as the active agents in the preparation of nutrient medium as many of them may ultimately contribute themselves to the nutrition of A. bisporus. The diversity of microflora according to growing farmhouse and fruiting body of Agaricus bisporus were investigated. The aerobic bacteria and Bacillus as longer of turning stage of compost pile were increased. And, thermophilic actinomycetes and fluorescent Pseudomonas sp. showed high density after the pasteurization stage. But Tricoderma sp. was decreased toward the end of turning stage of compost pile. Ten mushroom farms was selected to research of microflora of fruiting body of button mushroom. The microflora showed significant difference according to mushroom farms. The bacteria density was 0.4~41.6×105 cfu/ml and the fungus was 1.3~3.9×103 cfu/ml. But The microorganism density was not significant change for the storage periods. These isolates were classified into Chryseobacterium indologenes(6 strains), Pseudomonas agarici(5 strains), Sphingobacterium multivorum(2 strains), Flavobacterium anhuiense(2 strains), Microbacterium sp.(10 strains), Pseudomonas sp.(13 strains) on the basis of 16 rDNA analysis. The most dominants of these species were Chryseobacterium indologenes and Pseudomonas agarici.

The winter mushroom, Flammulina velutipes, is one of the major economical crops cultivated in Korea. The total production have steadily increased approximately 40,161 M/T in 2005 to 61,057 M/T in 2009. Several bacteria have been known as the causal agents of certain diseases of cultivated button mushroom(Agaricus bisporus) oyster mushroom(Pleurotus ostreatus) and winter mushroom(Flammulina velutipes). It is well known as bacterial diseases of the cultivated mushroom such as brown blotch, mummy disease, bacterial pit, bacterial rot, weeping disease, ginger blotch, and drippy gill. Black rot has been recognized as a major problem within the mushroom industry. Pseudomonas tolaasii has been shown to be associated with a black disorder of the caps and stipe of the mushroom. Recently, P. tolaasii was isolated from disease cultivated winter mushrooms grown in Korea. Its symptom appeared as dark brown and sunken lesions on the caps and stipes of affected mushrooms. Inoculation of bacterial isolates into mushroom caps and stipes showed characteristic black rot symptoms and sunken lesions. Results of Gram stain, staining of flagella and biochemical tests identified these isolates as P. tolaasii. This was confirmed by pathogenicity, physiological and biochemical characteristics, and results of an analysis of the 16S rRNA gene sequences.

This study was carried out to decrease the royalties through the dissemination of domestic mushroom varieties by following UPOV (Union Internationale Pour la Protaection des Obtentions Vegetablue: the international union for the protection of new varieties of plants). Although RDA has bred various mushrooms of 25 species of 78 varieties since 1990, most of these were not cultivated in mushroom farms due to two reasons. One is the weaknees of spawn supply system caused by increasing own spawn production and the other is the difficulty in replacing a new variety due to the year-round production in bottle cultivation. Therefore, we made a program to disseminate domestic mushroom varieties. In 2010, five species of 14 varieties were provided as a form of spawn for 43 farms that do not have their own cultivation facility and 11 species of 18 varieties were provided for 45 farms that were equipped with their own cultivation facility. In addition, the breeders explained the cultural characteristics of their own varieties and consulted cultural farms. Since these activities were helpful to mushroom farmers, we plan to open the presentation for mushroom instructor in February 2011 with the consideration of the opinion of mushroom farmers to fully disseminate the spawn of domestic mushroom varieties for the bottle cultivation farms.

This study was carried out investigate to availability of commercial microbial pesticide and antibacterial activity of isolates isolated from different mushroom media. Ten commercial microbial pesticide and EM liquid were collected from different company. EM of these one has been used for control of mushroom disease and growth promotion at the mushroom farms. The density of bacteria and yeast in EM cultural liquid was higher than those of the crude liquid. The pH values of EM showed the low acid levels(pH 3.5~3.9) by organic acid secreted from microorganism. The kinds of organic acid was acetic acid, lactic acid and succinic acid. The dominant bacteria isolated from EM liquid was Lactobactillus sp.(21 strains), Acetobacter sp.(9 strains), PaeniBacillus sp.(9 strains) and others(12 strains). The organic acid bacteria isolated from fermentation foods( was 92 strains and the dominant genus was Weissella sp.(41 strains), Leuconostoc sp.(21 strains), Enterococcus sp.(9 strains), Lactococcus sp.(9 strains) and others(12 strains). And we isolated 2,500 bacteria from oyster mushroom and button mushroom cultural media for selection of antagonistic bacteria. Thirty five strains of these isolates showed very strong antagonistic activity. These strains were identified Bacillus amyloliquefaciens, Bacillus subtilis, Brevibacterium halotolerans, Pseudomonas libanensis, Stenotrophomonas maltophilia and Alcaligenes faecalis by 16S rDNA analysis.

To understand the saccharide contents and quantity of winter mushroom (Flammulina velutipes) according to its growth temperature, we measured the saccharide contents at different growth temperature. In our results, the saccharide of its fruiting body turned out to be mainly composed of xylose, trehalose and mannitol in all treatments. In the other hand, Ribose, myo-inositol and sucrose were detected in some treatments. The quantity of trehalose decreased as the growth temperature increased with a variation of its quantity depending on the isolates used in the experiments. In the case of xylose and mannitol, detected in all treatments, the pattern of their quantities was not possible to be profiled and the pattern might be largely depending on the isolates. However, the quantities of xylose and mannitol were largely in a direct proportion and the fluctuation of their quantities was congruent with the exception of ASI 4103, ASI 4166 and ASI 4065. The xylose quantity of ASI 4103 and ASI 4166 increased until the temperature was up to 10℃ and decreased when the temperature was above 10℃. That of ASI 4065 decreased as the temperature rose and increased when above 13℃. The mannitol quantity of ASI　4065 and ASI 4166 decreased until the temperature was up to 10℃ and increased when the temperature was above 10℃. That of ASI 4103 increased as the temperature rose and decreased when above 13℃.

We investigated 18 amino acid contents in the fruiting bodies of Agaricus strains to understand their variation in the harvesting periods. Total content of amino acid was different depending on the isolates. In the case of A. bitorquis and A. brunnescens, the total amino acid content increased and correlated with the harvesting periods. For commercial strains of A. bisporus, such as Yangsongi-505, Yangsongi-705 and commercial strain A collected in the market, it increased until the second harvesting period, but decreased in the third harvesting period indicating that these three isolates exhibit the exceptions for the general trend. Among isolates we used in our experiments, the isolate of A. brunnescens contained the highest and two isolates of A. bisporus were the lowest in total amino acids. We found two trends for the contents of each amino acid in the fruiting bodies of Agaricus spp.. One is the same with the trend of the total amino acid content and the amino acid content were largely correlated with the harvesting period. The other is the trend of increase of amino acid until the second harvesting period and decrease of amino acid in the third harvesting period, which was also shown in the total amino acid content of A. bisporus. Generally, the most amino acid contained in Agaricus was cysteine and followed by phenylalanine, glutamic acid, lysine, proline and histidine.

This study was conducted to evaluate the feeding value of the spent mushroom (Pleurotus eryngii) substrates (SMS) in laying hens (Hy-Line Brown). The fresh spent mushroom (Pleurotus eryngii) substrates collected from the DOJUN farm were fermented with Bacillus subtilis EJ3 for 2 weeks. A total of twenty-four laying hens were fed corn-soy based experimental diets containing 0% (control), 5% (T1), 10% (T2) and 15%(T3) fermented SMS for 7 weeks. There were no significant differences among the treatments in egg production, egg weight, egg mass, feed conversion and viability during the experimental period. Feed intake was significantly lowered in control (118.3 g) than T1 (121.9 g), T2 (120.3 g) and T3 (122.4 g). There were no significant differences among the treatments eggshell breaking strength, thickness and haugh unit, whereas the yolk color of T1, T2 and T3 were significantly heavy than T0. The palatability of boiled meat was significantly better in the T3 laying hens than in the T0 laying hens. In conclusion, fermented SMS can be used as resource of feed in laying hen feed at 5.0-15% level without effect on performance and egg qualify.

The mannanase, cellulase and xylanase-producing thermophilic bacteria, designated EJ3, was isolated from fresh spent mushroom (Pleurotus eryngii) substrates taken from the DOJUN farm located in Keyongnam, Korea. The isolate EJ3 was facultatively anaerobic and grew at temperature ranging from 20℃ to 60℃ with an optimal temperature of 37℃. The DNA G+C content of the isolate EJ3 was 45 mlo%. The major fatty acids were anteiso-15:0 (38.9%), 17:0 (7.4%), and iso-15:0 (38.2%). The 16S rRNA gene sequence similarity between the isolate EJ3 and other Bacillus strains varied from 97% to 99%. In the phylogenetic analysis based on these sequences, the isolate EJ3 and Bacillus subtilis clustered within a group and separated from other species of Bacillus. Based on the physiological and molecular properties, the isolate EJ3 was classified within the genus Bacillus as Bacillus subtilis EJ3. The optimal growth condition of B. subtilis EJ3 were pH 6.0 and 55℃, respectively.

We investigated 18 amino acid contents in the fruiting bodies of Agaricus strains to understand their variation in different growth temperature. Although total content of amino acid was more or less different depending on the isolates, it increased until the temperature was 22℃ and decreased above 22℃. Commercial strains of A. bisporus, such as Yangsongi-505, Yangsongi-705 and commercial strain A collected in the market exhibited these general trend in total amino acid content. However, another commercial strain B did not follow these trend. A strain of A. bitorquis did not produce mushrooms at 10 and 13℃, but mushrooms harvested in the other treated temperatures exhibited with the similar patten. Generally, the most amino acid contained in Agaricus spp. was cysteine and followed by phenylalanine, glutamic acid, lysine, proline and histidine. We detected two trends for the contents of each amino acid in the fruiting bodies of Agaricus spp. grown at the different growing temperature. Some amino acid contents were largely correlated with the growth temperature, but the others showed the trend of increase until 22℃ and decrease above 22℃.

[Background] Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3’-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. However, the molecular mechanisms of inflammatory mediator’s activity by cordycepin remain poorly understood. In the present study, we investigat-ed the effects of cordycepin on the anti- inflammation cascades in lipopolysaccharide (LPS)-stimulated BV2 microglia cells. [Methods] Cordycepin were administered and their effects on LPS-induced pro-inflammatory mediators and MAP kinases were monitored by Western blotting and RT-PCR analysis. [Result] Cordycepin significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), and pro- inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in a concentration- dependent manner without causing cytotoxicity. Also, cordycepin suppressed inducible NO, synthase (iNOS) and cyclooxygenase-2 (COX-2) expression on the mRNA and protein level. In addition, cordycepin suppressed NF-κB translocation by blocking IkappaB- α (IκB-α) degradation and inhibited the phosphorylation of Akt, ERK-1/2, JNK, and p38 kinase. Our results indicate that the inhibitory effect of cordycepin on LPS -stimulated inflammatory mediator production in BV2 microglia is associated with the suppression of the NF-κB, Akt, and MAPK signaling pathways. Conclusion: Anti-inflammatory properties of cordycepin may be useful for treating the inflammatory and deleterious effects of microglial activation in response to LPS stimulation.

We have purified and characterized of metalloprotease metalloprotease from Nomuraea atypicola. N. atypicola was cultured in Sabouraud medium supplemented with powdered pupae. The metalloprotease from culture supernatant was purified to electrophoretically homogeneous state. The molecular mass of metalloprotease from N. atypicola was 50 kDa. The enzyme was most active at pH 8.5 and 40oC and stable at pH 5.0-7.0 and up to 40oC. The activity was inhibited by o-phenanthroline and EDTA. The N-terminal amino acid sequence of the enzyme showed a similarity to those of proteases (Metallo peptidase M36 family (Fungalysin)) from Coccidioides posadasii and Aspergillus fumigatus. The enzyme was found to be Fungalysin-like metalloprotease. cDNA encoding metalloprotease from N. atypicola was amplified by PCR using oligonucleotides deduced from the N-terminal endo peptide sequence, 5’- and 3’-RACE. Predicted enzyme structure consists of 637 amino acids with pro- and signal sequences. The mature enzyme had 391 amino acids and its deduced amino acid sequence coincided completely with the N- terminal amino end (20 amino acids) of metalloprotease purified from N. atypicola. We are studying on expression of the metalloprotease gene in Escherichia coli.

[Introduction] Mushroom constituents have been found to be highly effective in the prevention and treatment of lifestyle diseases such as lipidosis, high blood pressure, and diabetes, which are closely linked to eating habits, and several varieties of functional foods have been developed from these constituents. As a result, doctors of Western medicine in particular, who in the past have been dismissive of herbal or Oriental medicine, are now tending to take a more proactive stance toward adopting the better aspects of alternative, complementary or traditional medicine. Natural remedies and folk medicines have been incorporated into the treatment of cancer, for example, and in the same way it has become common to incorporate mushrooms into treatment regimes before and after surgical interventions and alongside therapies such as chemotherapy, radiation therapy, and hormone therapy. Mushroom chitosan, which is investigated in this study, is a novel functional ingredient made from the mushroom Flammulina velutipes (Curt.:Fr.) Sing., which is the most common edible mushroom to be artificially cultivated in Japan and has long been part of the Japanese diet. Unlike chitosans of crustacean origin, mushroom chitosan is rich in the main structural component β-glucan, and this dietary fiber is expected to have positive functions within the body. In particular, there are hopes that mushroom chitosan will inhibit cholesterol and fat absorption in the small intestine, and suppress total cholesterol and neutral fat levels in the serum. Previous human trials have confirmed the anti-metabolic syndrome efficacy of supplements containing mushroom chitosan. Here we report the ameliorative effects of mushroom chitosan in an animal model of genetic obesity and lipidosis. [Methods] Mushroom chitosan (RSK2, Ricom Corporation) was administered at different doses to Crj:(ZUC)-fa/fa rats an animal model of obesity and hyperlipidemia continuously for 10 weeks. The rats were kept at a temperature of 22±1ºC and humidity of 60±10%, and illuminated with fluorescent lamps for 12 h/day (07:00~19:00). Body weight, food consumption, body condition, and hematological and blood biochemical blood parameters were measured, and pathological examination (pathological analysis of hepatic lipid droplets) was performed using HE staining. [Results and discussion] Mushroom chitosan (RSK2) showed high efficacy in suppressing weight gain in Crj:(ZUC)-fa/fa rats presenting obesity due to genetic lipidosis. The smallest effective dose was 3 mg/kg. In addition, values for neutral fat, β-lipoproteins and total lipid due to ingestion improved to the normal values. Moreover, the pathological study of the liver revealed a decrease in lipid droplets appearing in the central zone of the hepatic lobule and a decrease in fat deposition in the liver in the group that ingested mushroom chitosan. The results also suggested that serum lipid levels were improved through egestion of excess fat with the feces. Mushroom chitosan (RSK2) was shown to be effective in controlling increase in serum lipids as it has lipase-inhibiting activity, and was shown to control fat deposition in internal organs through egestion of excess blood lipids with the feces. Mushroom chitosan is thus a functional food that is effective in preventing and treating contemporary lifestyle diseases.

[Background] Cordyceps militaris is a traditional popular mushroom, produces an important bioactive compound Cordycepin (3’-deoxyadenosine) used for the tonic and medicinal purpose in eastern Asia. Cordycepin is reported to possess many pharmacological activities including immunologically stimulating, anti-tumor, anti-virus, and anti-infection effects. [Methods] Growth inhibition of human leukemia cells was assessed by MTT assays. The determination of apoptotic cell death was performed by flow cytometry analysis, agarose gel electrophoresis and DAPI fluorescent staining methods. The apoptotic-regulated gene markers in both death receptor- and mitochondria-mediated apoptotic pathways were detected by RT-PCR and Western blot analysis etc. [Results] It was found that inhibition of cell proliferation was observed for human leukemia U937 and THP-1 cells treated with cordycepin in a dose-dependent manner. Cordycepin induced morphological change and apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptotic-sub G1 phase. Apoptosis of U937 and THP-1 cells by cordycepin was associated with a down-regulation of anti-apoptotic Bcl-2 and inhibitor of apoptosis proteins (IAPs) expression. Cordycepin treatment induced the proteolytic activation of caspase-3, caspase-8 and caspase-9, and a concomitant inhibition of poly(ADP-ribose) polymerase (PARP), β-catenin and phospholipase (PLC)-γ1 protein. Conclusions: Our results indicated that the apoptotic processes caused by cordycepin are mediated by the regulation of the Bcl-2 and caspase family in human leukemia U937 and THP-1 cells. Our data also suggested that cordycepin may be a potential chemotherapeutic agent for the treatment of leukemia cancer patients.

In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti- inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, β-glucan, and polyphenol contents were also measured. The β-glucan and polyphenol contents were the highest in Pleurotus cornucopiae var. citrinopileatus (yellow Pleurotus) of all species. The yellow Pleurotus also exhibited the highest physiological activity as assessed by the DPPH IC, Cytotoxic activity, ACE activity and so on. To confirm the mechanism for the physiological activity of the yellow Pleurotus, we performed further examinations within ICR mice. The yellow Pleurotus reduced glucose, triglyceride and total cholesterol levels in the ICR mice blood for 4 weeks after feeding, and also significantly lowered both GOT and GPT levels. Taken together, our data indicates the yellow pleurotus is a promising functional food ingredient.

To develop new nutraceuticals from mushrooms, nutritional properties of two kinds of Hypsizygus marmoreus fruiting bodies were investigated. Hypsizygus marmoreus (white) contained 21.63% H. marmoreus (brown) contained 27.34% of crude protein, 55.75% of total sugar, and 2.69% crude lipid and 7.77% ash. It also contained 10.04 mg% Na, 19.28 mg% Ca, 9.30 mg% Fe and its calorie value was 356.57 Kcal per 100 g. Free sugar and amino acid content of H. marmoreus (brown) were determined.

To determine the optimal media conditions for the detection of the cellulolytic activity in Ganoderma neo- japonicum, we varied three media conditions: dye reagent, pH, and temperature. First, we evaluated the use of four dyes, Congo Red, Phenol Red, Remazol Brilliant Blue, and Trypan Blue. To observe the effect of pH on various media spanning acidic and alkaline pHs, ranging from 4.5 to 8.0. Furthermore, in order to research the effect of temperature on the clear zone and the fungus growing zone, we tested temperatures ranging from 15 to 35℃. On the whole, the best protocol called for Ganoderma neo-japonicum transfer onto media containing Congo red with pH adjusted to 7.0, followed by incubation at 25℃ for 5 days. Our results will be useful to researchers who aim to study extracellular enzyme activity in Ganoderma neo-japonicum.

This study was carried out to investigate the effects of Hericium erinaceus extract on the Anti- inflammatory and Anti-bacterial. To investigate in vitro Anti-bacterial activity assay, hot water extracts of Hericium erinaceus carried out using a broth micro-dillution method following the Clinical and Laboratory Standards Institute guidlines(Citron and Hecht, 2003; Jorgensen and Turnidge, 2003). In the next experiment, to investigate anti-inflammatory test, the RAW 264.7 macrophage, cells was cultured using DMEM including the 10% FBS. Hericium erinaceus extract has the effects of anti-inflammation and, which may be due to its inhibitory potential on the macrophage activation. Furthermore, Hericium erinaceus extract has the anti-bacterial effects through the inhibitory Propionibacterium acnes(P. acnes) and periodontopathic bacteria, Porphyromonas gingivalis , Fusobacterium nucleatum and Tannerella forsythia. The above results suggest that Hericium erinaceus hot water extract could be applicable for improvement of several inflammatory disease.

Ganoderma lucidum has long been used as a potent medicinal plant. In addition, recent studies showed that G. lucidum is used to prevent or treat various human diseases such as allergy, hepatopathy, hypertension, cancer and diabetes. In this study showed that ethanol extract from G. lucidum have potential inhibitory effects of glycerol 3-phosphat acyltransferase (GPAT) activity and increase of glucose uptake on skeletal muscle cells. Inhibition of GPAT, which catalyze the first step in de novo TAG synthesis, has been proposed as one of the drug targets for insulin resistance and type-2 diabetes. G. lucidum extract inhibited GPAT activity. Bioassay-guided fractionation and isolation of bio-active substances from ethanol extracts of G. lucidum were carried out by using chromatographic techniques and in vitro GPAT enzyme assay. One of bioactive molecules, ergosterol peroxide, was isolated and identified by physical and spectral properties. In this study showed that ethanol extract from G. lucidum and ergosterol peroxide have potential anti- diabetes effects through the increase of glucose uptake. This was as sociated with increased activity AMP-activated protein kinase(AMPK). AMPK is another regulatory protein in the glucose uptake pathway and energy metabolism. The result was that the increase of glucose uptake by G. lucidum extract might be mediated by AMPK.

Mushrooms including Mycoleptodonoides aitchisonii are used as foods and employed as folk remedies for diabetes and inflammatroy diseases. This study analyzed anti-diabetic effects of Mycoleptodonoides aitchisonii, which grows in some areas such as Gangwon-do and Jeju-do in Korea, as insulin-derived phosphorylation. When 100 ng/ml of IL-6, inflammatory cytokine was given to SK-hep1, HepG2, Akt phosphorylation by insulin was found to be remarkably reduced. In addition, metformin, Antidiabetics serving as positive control in liver was used. Mycoleptodonoides aitchisonii used for analyzing anti-diabetic effects in this test didn’t give a great impact on the decrease in Akt phosphorylation by IL-6 at high concentration. However, fruit body in Mycoleptodonoides aitchisonii inhibited the decrease in Akt phosphorylation by IL-6 according to the concentration. At the highest concentration 100㎍/ml, it had an effect of increasing Akt phosphorylation to 77%, which was decreased by 50% compared to the insulin treated group by IL-6. Therefore, Mycoleptodonoides aitchisonii fruit body used in this test activated Akt phosphorylation inhibited by IL-6 and showed a possibility of significant anti-diabetic effects compared to positive control(metformin).

Neurotrophic factors are essential to maintain and organize neurons functionally; thereby neurotrophic factor-like substances or their inducers are expected to be applied to the treatment of neurodegenerative diseases such as stroke. In the present study, we firstly examined the effects of ethanol extracts of Hericium erinaceus (HE, Yamabushitake), on nerve growth factor expression in neuronal cells. HE extract promoted NGF expression in a brain tissue. Here we assessed neuroprotective effects of HE with a transient global cerebral ischemia model. Global cerebral ischemia was induced by occluding both common carotid arteries. Treatment with HE was initiated after ischemia induction and given once a day for 7 consecutive days. Neuronal cell loss in CA1 of hippocampus was significantly decreased and the performance in the Morris water maze was significantly improved in rats administered HE. We conclude that treatment with HE attenuated learning and memory deficits, motor functional disability, and neuronal cell loss induced by global cerebral ischemia. These results suggest that HE may be a potential candidate for the treatment of vascular dementia.

Pleurotus eryngii (also known as king trumpet mushroom, french horn mushroom, king oyster mushroom) is an edible mushroom native to Mediterranean regions of Europe, the Middle East, and North Africa, but also grown in parts of Asia. It has the ability to produce various biologically active compounds and possesses a well-developed ligninolytic enzyme system that participates in the degradation of lignin and different aromatic compounds. In this study, we investigated the protective effects of the ethyl acetate extract of Pleurotus eryngii (PEE) on the development of atopic dermatitis (AD) in human keratinocyte HaCaT cells. Keratinocytes, one of major cell types in the skin, can be induced by TNF-α and IFN-γ to express thymus and activation-regulated chemokine (TARC/CCL17), which is considered to be a pivotal mediator in the inflammatory responses during the development of inflammatory skin diseases, such as AD. In addition, normal T-cell–expressed and secreted chemokine (RANTES) is a (C-C) chemokine released by T lymphocytes, other inflammatory cells, and platelets and plays an important role in allergic inflammatory processes. Pretreatment of HaCaT cells with PEE suppressed TNF-α/IFN-γ-induced protein and mRNA expression of CCL17 and RANTES. PEE significantly inhibited TNF-α/IFN-γ-induced NF-κB activation. These results suggest that PEE may exert anti-inflammatory responses by suppressing TNF-α and IFN-γ-induced activation of NF-κB in the keratinocytes and might be a useful tool in therapy of skin inflammatory diseases.

This study was conducted to investigate optimum processing procedure of fresh-cut button mushrooms. The experiments were done with different processing parameters including washing method, washing time, sanitation, and cutting time. Fresh mushrooms were washed by swirling water (SW), lift type-immersion washing (LIW), or combined LIW and water spray (LI+WS). Mushroom samples were washed by LIW for 0, 1, 3, and 5 minutes separately. Mushrooms were sanitized with 0, 50, or 100 ㎕L-1 chlorine solution (pH 7) for 60 seconds. Mushrooms were sliced at different times (before washing, after washing, or after drying). The combined washing treatment, LIW+WS was effective in maintaining better appearance and higher Lightness color value among treatments. Optimum washing time to remove foreign materials and maintain color was 3 minutes when mushrooms were washed by LIW. Samples sanitized with 50 ㎕L-1 chlorine reduced initial aerobic bacterial population and had only slight residual chlorine odor. Fresh-cut mushrooms sliced after washing had the lowest loss among samples. The optimized washing, sanitation, and cutting time parameters can be used for sequential processing of fresh-cut button mushrooms.

The Lentinula edodes is the third most widely produced mushroom in the world. It is a rich source of proteins, fats, carbohydrates, fiber, vitamins, minerals and β -glucan. The aim of the this study was to evaluate the physicochemical characteristics changes such as proximate composition, minerals, and contents of β -glucan according to the different product area. And we evaluated the antioxidant properties of L. edodes. Also, we determined the standardization of manufacture process for commercial products using L. edodes. Eleven L. edodes were cultivated in oak, and samples were collected by different product area. And the L. edodes cultivated in sawdust have been producted in Jangheung. All samples are shade dried and those dried samples were used for this test.

This study was conducted to determine changes in quality and microbial population of intact and fresh-cut button mushrooms (Agaricus bisporus). Freshly collected mushroom was cut into approximately 0.5 cm thick slices and washed in tap water or 100 ㎕L-1 chlorine solution (pH 7) for 60 seconds. Intact mushrooms were washed in the same condition as cut samples. Both intact and fresh-cut samples were then dried, packaged in 50㎛ poly ethylene bags, and stored at 5 ℃ for up to 9 days. Quality and microbial safety parameters such as gas composition, color, off-odor, visual quality, electrical conductivity, E. coli / coliform count, and total aerobic population were evaluated during storage. All sample packages exhibited a rapid depletion of O2 (to ~0 kPa) and accumulation of CO2 (10.3 to 12.6 kPa) throughout the storage period. No significant color difference was found between tap water and chlorine. However, chlorine treatment was effective in reducing off-odor development of intact and fresh-cut samples at the end of storage. The chlorine application also reduced aerobic bacterial populations. Both Intact and cut mushrooms had ≤ 5 log CFU/g of total aerobic plate counts until the end of storage. Fresh-cut samples regardless of chlorine sanitation had higher overall visual quality score than intact samples. Results indicated that fresh-cut mushrooms treated with chlorine maintained quality and shelf-life throughout the 9 day storage period.

Many Wild Mushrooms were collected at Yuchi recreational Forests areas for 1 day in July 2010 and 3 days in August 2010. They were identified and examined with references. According to the result, Species diversities are 1 division, 2 classes, 3 subclasses, 5 orders, 13 familys, 20 genera and 28 species. Dominant famillie are Tricholomataceae. Dominant genus are Amanita. Dominant species are Amanita verna (Bull. : Fr.) Roques and Coriolus versicolor (L. : r.) Quel. Resources of Wild Mushrooms were 2 species in edibility, 3 species in clulture, 12 species in toxine, 5 species in medicine, 2 species in anticancer, 11 species in ectomycorrizahe and 4 species in rotten wood.

The genetic diversity of 16 Ganoderma strains was investigated by rDNA-ITS sequencing. Alignment analysis showed that whole length of internal transcribed spacer(ITS1+ITS2) was 500bp and with 139 variation sites (accounting for 23.3％, ITS1 was 66 and ITS2 was 73), 337 conserved sites (accounting for 72.2％), 59 informative sites (accounting for 9.88％), 86 conversion sites (G-A, C-T), 13 transversion site(C-G, T-A). The ratio of transition and transversion in ITS1 was higher than that in ITS2, and the variable sites of ITS2 were more than those of ITS1. The genetic distance among 16 Ganoderma strains is from 0 to 0.121. The genetic distance between G. lipsiense and F-1 was 0, and the genetic distance between Heizhi 02 and Huizhou, Jingda, G. capense was 0.121, 0.117 and 0.120, respectively. The 16 Ganoderma strains were classed into 4 groups. The biggest group is comprised of 12 strains, including Xinzhou, Huizhou, Jingda, 902, F-1, Xianzhi, Meiluo, Taishan, G. applanatum, 05, G. luteomarginatum. The G. atrum and G. sinense were clustered into one group. The G. capense and Zhongzhi was independent group, respectively. These results showed that there were some genetic difference among groups, and there was lower genetic diversity among strains in same groups.