• Journal of Life Science
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• v.22 no.6
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• pp.828-836
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• 2012

The objective of the current study was to determine the effects of the extracts isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants on mouse splenocytes, B cells, and macrophages in vitro. The ethanol extracts B (EEB) directly induced the proliferation of spleen cells in a dose-dependent manner and increased IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ synthesis. The EEB also increased the proliferation of B cells in a dose-dependent manner. The production of immunoglobulin M, G1, G2a, G2b, and IgG3 in the presence of the EEB increased progressively in the culture supernatant. When the EEB were used in macrophage cell line (RAW264.7) stimulation, there was a marked induction of NO synthesis in a dose-dependent manner and an increased IL-6, TNF-${\alpha}$, and GM-CSF synthesis. Intraperitoneal injection with EBB showed life prolongation effect of 16.1% in mice previously inoculated with sarcoma-180, respectively. These results suggest that the capacity of the EEB isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants seems to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, Flammulina velutipes EEB could be used as a biological response modifier having possible therapeutic effects against immunological disorders. This study also showed that functional components of Flammulina velutipes were possibly improved by incorporating oriental herbal plants in a growth medium.

• Journal of Mushroom
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• v.12 no.3
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• pp.193-200
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• 2014

Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium were analyzed for their proximate composition, protein-bound polysaccharide, nucleic acid and amino acids. The content of ash and crude fiber in F. velutipes fruit body were higher than F. mycelium and C. militaris mycelium. C. militaris mycelium showed the highest crude fat content while F. velutipes fruit body had lowest. Nitrogen free extract content of the samples varied from 56.8% in F. velutipes fruit body to 61.9% in F. velutipes mycelium. The compositions of total protein and total free sugars of protein-bound polysaccharide were found to be significant differences for all samples. Nucleic acid related compounds were identified the 5'-GMP, 5'-XMP, 5'-IMP in all samples. The content of total nucleic acids were high in the orders of F. velutipes myclial (286.71 mg%), F. velutipes fruit body(187.36 mg%) and C. militaris mycelial(76.85 mg%). The highest content of 5'-GMP was found in F. velutipes fruit body. The most nucleic acid of F. velutipes mycelial and C. militaris mycelial were the 5'-XMP. As for the analysis of total amino acids, seventeen amino acids were identified by HPLC and the major amino acid was glutamic acid in all samples. The content of total amino acids were high in the orders of F. velutipes fruit body(19,919 mg%), F. velutipes mycelium(19,018 mg%) abd C. militaris mycelium(18,965 mg%). We determined the developing new food product such as amino acid drink and amino acid containing food using extracts of Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium.

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.111-120
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• 1997

The characteristics of monosporous isolates of winter mushroom [Flammulina velutipes (Curt. ex Fr.)] were investigated to obtain useful breeding materials. Within monokaryons from cultivars which have a white colored fruitbody showed narrow genetic variation, while domestic strains which have a brown colored fruitbody showed wide variation. The mating type of the white strains was A1A2B1B2 genotype, but that of the domestic brown strains were A3A4B3B4. In intra-crossing, the genetic stability of dikaryons mated by monokaryons from white strain was less than that of parents. While in brown strain, dikaryons with high yield and color variation were obtained.

• The Korean Journal of Food And Nutrition
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• v.23 no.3
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• pp.376-380
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• 2010

Mushrooms(Pleurotus ostreatus, Agaricus bisporus and Flammulina velutipes) are popular food sources in Korea, and have been reported as therapeutic foods, useful for preventing various diseases. In this study we researched HPLC conditions for the determination of nucleic acids in extracts of the three type of mushrooms. The method for nucleic acids analysis of mushrooms was developed using HPLC with UV detection. To determine the nucleic acids, mushroom extracts were extracted in hot water at $90^{\circ}C$ by reflux extraction for 1 hr. Then, the extracts were hydrolyzed by enzymes RP-1G and 50000G. The HPLC conditions were simple, rapid, and sensitive, and were applicable for the analysis of 4 nucleosides(cytidine, uridine, guanosine and inosine) and 3 mono-nucleotides(5'-CMP, 5'-UMP, and 5'-IMP) in the mushrooms. The nucleic acids in the mushrooms were cytidine, guanosine, inosine, uridine, 5'-CMP, 5'-IMP, and 5'-UMP. The analysis results for total nucleic acids in the mushroom extracts(Pleurotus ostreatus, Agaricus bisporus, and Flammulina velutipes) indicated levels of 25.28, 27.75, and 19.87 mg/g, respectively. In conclusion, this method can be used successfully for qualitative and quantitative analysis of nucleic acids in Pleurotus ostreatus, Agaricus bisporus, and Flammulina velutipes.

• Journal of the Korean Wood Science and Technology
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• v.14 no.3
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• pp.36-42
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• 1986

The production media and the enzymatic charateristics of laccase from Flammulina velutipes were investigated. The activity of laccase during incubation reached to the maximum at the 40 days of incubation in the case of Barley straw medium. The maximum laccase activity in Barley straw medium was 5 and 16 times higher than those in Onion basic and Sawdust media, respectively. The laccase from Flammulina velutipes has the optimum pH of 6.6 and showed to be stable at relatively broad pH range. 4.5-9.5. Temperature stability showed that above 96% activity could be preserved after holding at 40$^{\circ}C$ for 40 minutes. At the above 70$^{\circ}C$, the laccase activity decreased very rapidly. The Km value of the laccase was estimated to be 28.0 mM which is much higher than that of the laccase from Pleurotus ostreatus. Organic solvents for precipitiation of the enzyme did not inactivation the laccase. Sodium azide which was added for preventing microbial deterioration affected significantly the inactivation of laccase, but this activity was recovered completely by precipitating the enzyme with acetone.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.355-358
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• 2001

Selection of optima] nutrient sources and cultural methods for liquid spawn culture of Flammulina velutipes were carried out. The optimal temperature and pH for mycelial growth of F. velutipes were $20^{\circ}C$ and 6.0 to 7.5. respectively. In the 250ml ${Delta}$-flask culture. the amount of inoculum and culture period for the optimal mycelial growth of F. velutipes were 3 mycelial disks(diametcr 6mm) and 6 days, respectively. For the mass production of submerged culture. the optimal inoculum amount and aeration rate of F. velutipes were 5%(inoculum vol/medium vol.) and l.0vvm(vol of air/vol. of medium/min), respectively.

• Mycobiology
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• v.41 no.1
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• pp.37-41
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• 2013

Laccases (EC 1.10.3.2) are copper-containing polyphenol oxidases found in white-rot fungi. Here, we report the cloning and analysis of the nucleotide sequence of a new laccase gene, fvlac7, based on the genomic sequence of Flammulina velutipes. A primer set was designed from the putative mRNA that was aligned to the genomic DNA of F. velutipes. A cDNA fragment approximately 1.6-kb long was then amplified by reverse transcriptase-PCR using total RNA, which was subsequently cloned and sequenced. The cDNA sequence of fvlac7 was then compared to that of the genomic DNA, and 16 introns were found in the genomic DNA sequence. The fvlac7 protein, which consists of 538 amino acids, showed only 42~51% identity with 12 different mushroom species containing two laccases of F. velutipes, suggesting the fvlac7 is a novel laccase gene. The first 25 amino acids of Fvlac7 correspond to a predicted signal sequence, four copper-binding sites, and four N-glycosylation sites. Fvlac7 cDNA was heterologously overexpressed in an Escherichia coli system with an approximate expected molecular weight of 60 kDa.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.235-238
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• 2011

Agrobacterium harboring vector pCHBs with hygromycin phosphotransferase(hph) and hepatitis B virus surface antigen(HBsAg)gene was transformed into the mycelial culture of Flammulina velutipes. In particular, mild NaOH solution was treated to the mycelia before Agrobacterium infection step. This was purposed to generate putative surface wounds in the mycelial cell walls. The results showed that hygromycin-resistant($hyg^r$) mycelia could be obtained only from NaOH-treated mycelia but not from intact mycelia. The integration of $hyg^r$ gene in fungal genome was confirmed by PCR. In addition, a single transgene integration and heterologous protein expression in F. velutipes could be verified by Southern blot hybridization and western blot analysis, respectively. This study demonstrated an efficient tool for the Agrobacterium-mediated transformation of F. velutipes mycelia.

• Journal of Mushroom
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• v.7 no.1
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• pp.22-26
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• 2009

These experiments were conducted to find the optimum mixing rate of used media for saving the production cost of Flammulina velutipes. The materials for media formation was used needle-leaf tree sawdust, media that finished 1th cultivation of F. velutipes, corncob meal, and rice bran, wheat bran as nutrition source. We inoculated the F. velutipes in 14 kinds of different media types and checked the spawn growth speed, fruit body quality and quantity. Two nutrition agents, which is rice bran and wheat bran, did not affected the incubation period, but the effective stem number, quality and quantity of fruit body was better at rice bran than wheat bran. The quality of fruit body produced at mixed 20% of used media (needle-leaf tree sawdust 60% + used media 20% + rice bran 20%) was similar to control plot (needle-leaf tree sawdust 80% + rice bran 20%), but the yield was improved 10% than control plot 130g. According as the used-media mixing amount increases, quality and quantity of fruit body became low remarkably. Therefore, the optimum mixing amount of used-media was 20% and it increased 10% of fruitbody yield.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.306-309
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• 1993

Vegetable oils supplemented to the basal medicum stimulated mycelial growth of Flammulina velutipes. The mycelial yield was increased 3.5 folds by addition of 3% (v/v) ricebran oil. Maximum mycelial yield (18.2mg/ml) was obtained by addition of 3.0% ricebran oil with 1.0% $CaCl_2$ to the basal medium. There was no significant difference between the liquid and solid spawn in the yield of sporophores.