• Journal of Society of Preventive Korean Medicine
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• v.13 no.2
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• pp.89-102
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• 2009

Objectives : The purpose of this study was evaluated hypoglycemic effect of culture broth of germanium-fortified Tricholoma matsutake mycelium and yeast. Methods : We examined $\alpha$-glucosidase inhibitory activity, blood glucose level, concentration of serum lipid, and serum metabolic variables of Tricholoma matsutake mycelium and yeast fortified Germanium. in streptozotocin induced diabetic rat. Results : In the $\alpha$-glucosidase inhibitory activity, germanium-fortified yeast was significantly higher than germanium-fortified Tricholoma matsutake mycelium. The hypoglycemic effects of germaniumfortified Tricholoma matsutake mycelium was higher than germanium-fortified yeast. The activity of alkaline phosphatase(ALP), aspartate aminotransferase(AST) and alanine aminotransferase(ALT) was significantly lower in the germanium-fortified Tricholoma matsutake mycelium and yeast than in diabetic control(DC) group and diabetic positive control(PC) group. The concentration of total cholesterol and triglyceride of germanium-fortified yeast was significantly lower than germanium-fortified Tricholoma matsutake mycelium, DC group and PC group. Conclusions : The results suggest that germanium-fortified Tricholoma matsutake mycelium and yeast have improvement effects in blood glucose, serum lipid and liver function.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.473-479
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• 2009

The purpose of this study was evaluated physiological activity effect of organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium and germanium-fortified yeast. Proliferation Tricholoma matsutake mycelium and yeast was inhibited by addition of germanium. Contents of organic germanium in Tricholoma matsutake mycelium and yeast was increased in dose-dependent manner. And low concentration(1,000 ppm) of germanium in mycelium was almost changed organic germanium. In the result of antioxidant activity as SOD-like activity, contents of total polyphenol compound and electron donating ability, activity of germanium-fortified Tricholoma matsutake mycelium was higher than that of germanium-fortified yeast. To evaluate of antitumor effects in vitro, we examined nitric oxide production of Raw 264,7 cell and cytotoxicity of HT1080 cell by MTT assay. Nitric oxide production of germanium-fortified Tricholoma matsutake mycelium was shown low level in low concentration(1,000 ppm) than other groups. The anticancer effect of germanium-fortified Tricholoma matsutake mycelium on HT 1080 cell was indicated a strong inhibitory effect in low concentration(1,000 ppm). These results suggest that organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium has valuable physiological activities as antioxidant and anticancer effect, and it was higher than that of germanium-fortified yeast.

• Mycobiology
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• v.35 no.2
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• pp.54-61
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• 2007

Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.365-370
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• 2008

This study was performed to investigate the influence of extracts from mixed culture with Tricholoma matsutake mycelium and Cordyceps militaris mycelium on hypoglycemia in streptozotocin-induced diabetic rats. Experimental animals were divided into 7 groups : normal control group(NC), diabetes control group (DC), positive control group(PC), non-fermented OCM(oriental medicine & cereal medium) extracts group (UM), crude polysaccharide of non-fermented OCM extracts group (UME), fermented OCM extracts group (UF), crude polysaccharide of fermented OCM extracts group (UFE). NC, DC groups were orally administered saline, PC group was orally administered acarbose. UM, UME, UF, UFE groups were orally administered each extract once a day for 14days. Blood glucose level was lower in the all administering OCM extract groups (UM, UME, UF, UFE) than in the diabetes group(p<0.05), and specially UF, UFE groups were similar to tendency of PC group. ALT, ALP activity in OCM groups were not significantly lowered than PC group(p<0.05). AST activity was not different with PC group. The results of this study show that extracts from mixed culture with Tricholoma matsutake mycelium and Cordyceps militaris mycelium may have a beneficial effect on the hypoglycemia.

• KSBB Journal
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• v.18 no.1
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• pp.45-50
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• 2003

In this study, the optimum conditions for mycelium culture of the mushroom Tricholoma matsutake and the inhibitory effect of the mycelium extracts no tyrosinase activity have been examined. When the extracts of the Tricholoma matsutake mycelia were tested for inhibitory activity on tyrosinase, it was found that the components extracted with ethyl acetate and water showed the highest inhibitory activity. The effect of antioxidants on the growth of mycelium and tyrosinase-inhibiting activity was also investigated. The results showed that tocopherol inhibited the growth in a concentration-dependent manner. In terms of tyrosinase-inhibiting activity, however, tocopherol was found to enhance the inhibitory activity.

• Journal of the Korean Wood Science and Technology
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• v.35 no.5
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• pp.109-117
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• 2007

Methyl trans-cinnamate is a significant flavor compound of Tricholoma matsutake. Attempts were made to produce this compound by culturing the mycelium using submerged culture. No methyl trans-cinnamate could be detected when the mycelium was cultured in the basal liquid medium. However, the addition of Pinus desiflora extracts to the medium, methyl trans-cinnamate was largely produced. To find out compounds or fractions inducing methyl trans-cinnamate, dichloromethane fraction obtained from the wood extracts of P. densiflora was subjected to column chromatography. Three sub-fractions were obtained from the $CH_2CI_2$ fraction. Submerged cultured mycelium treated with sub-fraction 1 has the highest content of methyl trans-cinnamate. Maximum methyl trans-cinnamate $(470.2{\mu}g/g)$ was obtained when the first sub-fraction of dichloromethane fraction of the P. densiflora wood extracts was added to the medium. This indicates that wood extracts of P. densiflora contains inducer of the methyl trans-cinnamate production in the T. matsutake submerged culture.

• Mycobiology
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• v.51 no.5
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• pp.354-359
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• 2023

Tricholoma matsutake has been the most valuable ectomycorrhizal fungi in Asia because of its unique flavor and taste. However, due to the difficulty of artificial cultivation, the cultivation of T. matsutake has relied on natural growth in forests. To cultivate the T. matsutake artificially, microorganisms in fairy rings were introduced. In this study, we isolated 30 fungal species of microfungi from the soil of fairy rings. Among them, one single fungal strain showed a promoting effect on the growth of T. matsutake. The growth effect was confirmed by measuring the growth area of T. matsutake and enzyme activities including a-amylase, cellulase, and b-glucosidase. In comparison with control, microfungal metabolite increased the growth area of T. matsutake by 213% and the enzyme activity of T. matsutake by 110-200%. The isolated fungal strain was identified as Penicillium citreonigrum by BLAST on the NCBI database. The Discovery of this microfungal strain is expected to contribute to artificial cultivation of T. matsutake.

• Mycobiology
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• v.29 no.4
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• pp.183-189
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• 2001

Extracellular amylase properties were examined with the mycelium of Tricholoma matsutake isolated from ectomycorrhizal roots of Pinus densiflora. The molecular weights of $\alpha$-amylase and glucoamylase were estimated as 34.2 kD and 11.5 kD, respectively, after eluted through Superdex 75 column. The optimum pH of the purified enzyme was found in a range of pH $5.0{\sim}6.0$, with a peak at pH 5.0. The activities of these enzymes were stable from $4^{\circ}C\;to\;30^{\circ}C$. The $\alpha$-amylase of T. matsutake readily hydrolyzed soluble starch and amylose-B, while it weakly hydrolyzed glycogen, dextrin, amylose and amylose-A. The main products of hydrolysis were confirmed to be glucose, maltose and maltotriose on the basis of the similarities in the thin layer chromatographic mobility.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1055-1059
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• 2013

The ectomycorrhizal fungus Tricholoma matsutake grows symbiotically with Pinus densiflora. Phenylalanine ammonia-lyase (E.C. 4.3.1.24) catalyzes the conversion of L-phenylalanine to trans-cinnamic acid. The role of fungal phenylalanine ammonia-lyase, however, has not been clear until now. In this study, the gene encoding phenylalanine ammonia-lyase (PAL), which was isolated from T. matsutake, was cloned and characterized. The PAL gene (tmpal) consists of 2,160 nucleotides, coding for a polypeptide containing 719 amino acid residues. The deduced amino acid sequence of tmpal from T. matsutake shows high identity (70%) with that from Laccaria bicolor. Comparative analysis of the PAL genes among T. matsutake and other species of the class Agaricomycetes showed that both active sites and binding sites were significantly conserved among these genes. The transcriptional analysis of the PAL gene revealed a differential gene expression pattern depending on the developmental stages (mycelium, primordium, stipe, pileus, and gills) of T. matsutake. These results suggest that the PAL gene in T. matsutake plays an important role in multiple physiological functions.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.89-93
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• 2003

In order to select suitable bioreactor type inhibiting cell stress during submerged culture of Tricholoma matsutake mycelium, the growth characteristics and ergosterol contents were investigated using the external-loop type of air-lift bioreactor (ETAB), balloon type of air bubble bioreactor (BTBB) and column type of air bubble bioreactor (CTBB). Dry weights of the T. matsutake in the BTBB, ETAB and CTBB were 12 g, 11.4 g, and 9.5 g per 1 litter, respectively. BTBB, ETAB and CTBB reached stagnant phases 16, 20, and 24 days after cultivation, respectively, The BTBB was more suitable for liquid culture of T. matsutake mycelium compared to other bioreactors owing to much mycelia product and short culture period. The ergosterol contents produced by the mycelium in the bioreactors were in sequence of BTBB, CTBB, and ETAB at every growth phase. BTBB might affect the mycelium on producing the smallest size of pellets. BTBB and CTBB got the mycelium precipitated and coagulated under operation of bioreactor sparser, whereas ETAB shown no effect of above phenomenon. A renovated bioreactor combined between a balloon shape of BTBB and an external-loop of ETAB was developed to enhance the efficiency of culture technique.