• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.355-358
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• 2001

팽나무버섯균의 배양환경조사에서 최적배양온도는 $20^{\circ}C$, pH는 $6.0{\sim}7.5$ 탄소원은 yellow sugar. potato starch. 질소원은 soybean flour. Yeast extract. 무기염류는 $KH_2PO_4,\;MgSO_4,\;7H_2O$를 0.05%(w/v)에서 우수한 생장을 나타냈다. 액체종균 배양 실험에서는 전 배양시 erlenmeyer flask에서 6 일간 $21^{\circ}C$, 125rpm으로 배양 후 본 배양시 5% 접종량, 1.0vvm 통기로 4-5 일 배양하였을 때 균체량이 높게 나타났다.

• 한국균학회지
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• 제21권3호
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• pp.212-216
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• 1993

팽나무버섯 재배에 있어서 각종 임산 및 농산폐기물을 기질로 사용한 결과, 나왕, 미송, 혼합톱밥, 커피박 및 땅콩박 중에서 커피박이 가장 자실체 생산이 양호하였으며 영양첨가제로 사용된 미강, 맥주박, 옥수수박, 대두박 및 채종박 중에서는 옥수수박에서 자실체 생산량이 높았다. 커피박과 옥수수박의 4 : 1 혼합비에서 자실체 생산이 129.38 g/bottle로 최대를 나타내었다.

• Mycobiology
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• 제32권1호
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• pp.11-16
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• 2004

Winter mushroom, Flammulina velutipes, needs low temperature during its cultivation. To save on farm costs, especially during summer, a strain adaptable to a higher or elevated-temperature must be developed. At the start of breeding program, parental strains which could endure high temperature were obtained. Seuenty four dikaryotic strains were collected and divided into four groups according to the nature of temperature. They also had different fruiting temperature. Finally we selected three brown strains ASI 4048, 4057 and 4072, and collected their spores. These selected strains can germinate even at a high temperature of $32^{\circ}C$, which were dramatically higher than the other strains. Based on these results, the new white strain adapted to mid-temperature by backcross mating was developed. Molecular markers were applied to select white fruitbody producing strains without cultivation. They showed a specific band which co-segregated with brown fruitbody forming strains in $BC_1F_1$ progenies. Selected white strains were tested under several elevated temperature conditions.

• 한국미생물·생명공학회지
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• 제46권3호
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• pp.300-312
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• 2018

Whole-genome sequencing of the wood-rotting fungus, Flammulina fennae, was carried out to identify carbohydrate-active enzymes (CAZymes). De novo genome assembly (31 kmer) of short reads by next-generation sequencing revealed a total genome length of 32,423,623 base pairs (39% GC). A total of 11,591 gene models in the assembled genome sequence of F. fennae were predicted by ab initio gene prediction using the AUGUSTUS tool. In a genome-wide comparison, 6,715 orthologous groups shared at least one gene with F. fennae and 10,667 (92%) of 11,591 genes for F. fennae proteins had orthologs among the Dikarya. Additionally, F. fennae contained 23 species-specific genes, of which 16 were paralogous. CAZyme identification and annotation revealed 513 CAZymes, including 82 auxiliary activities, 220 glycoside hydrolases, 85 glycosyltransferases, 20 polysaccharide lyases, 57 carbohydrate esterases, and 45 carbohydrate binding-modules in the F. fennae genome. The genome information of F. fennae increases the understanding of this basidiomycete fungus. CAZyme gene information will be useful for detailed studies of lignocellulosic biomass degradation for biotechnological and industrial applications.

• Journal of Microbiology and Biotechnology
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• 제19권7호
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• pp.681-684
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• 2009

This study was carried out to evaluate the inhibitory effect of Flammulina velutipes extracts on tyrosinase activity and to identify its biologically active component. The ethyl acetate and n-butanol extracts showed potent tyrosinase inhibitory activities. Subsequently, fractions of the n-butanol extract showed only a partial tyrosinase inhibitory activity. The most active compound of tyrosinase inhibitory activity was identified from the ethyl acetate extract as 1',3'-dilinolenoyl-2'-linoleoylglycerol (LnLLn) by comparing its mass, $^1H-$, and $^{13}C-NMR$spectral data with those previously reported in the literature. LnLLn showed tyrosinase inhibitory activity with an $IC_{50}$value of 16.1 ${\mu}g/ml$. These results suggest that the ethyl acetate extract of F. velutipes could be applicable for the development of a new whitening agent.

• 한국균학회지
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• 제17권1호
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• pp.35-38
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• 1989

팽나무버섯 균사체 DNA를 대장균 프라스미드인 PBR322를 이용하여 gene library를 작성 하였다. 팽나무버섯에서 ${\beta}-isopropyl$ malate dehydrogenase 유전자 클론을 얻었으며 이 클론은 대장균 Leucine 요구성 균주를 complementation 시켰다. 이 클론의 팽나무버섯 DNA 크기는 약 lKb였으며 BamH1과 Ava 1 제한효소 절단부위를 지니고 있었다.

• Mycobiology
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• 제38권4호
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• pp.331-335
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• 2010

In this study, we developed an efficient electroporation-mediated transformation system featuring Flammulina velutipes. The flammutoxin (ftx) gene of F. velutipes was isolated by reverse transcription-PCR. pFTXHg plasmid was constructed using the partial ftx gene (410 bp) along with the hygromycin B phosphotransferase gene (hygB) downstream of the glyceraldehydes-3-phosphate dehydrogenase (gpd) promoter. The plasmid was transformed into protoplasts of monokaryotic strain 4019-20 of F. velutipes by electroporation. High transformation efficiency was obtained with an electric-pulse of 1.25 kV/cm by using 177 transformants/${\mu}g$ of DNA in $1{\times}10^7$ protoplasts. PCR and Southern blot hybridization indicated that a single copy of the plasmid DNA was inserted at different locations in the F. velutipes genome by non-homologous recombination. Therefore, this transformation system could be used as a useful tool for gene function analysis of F. velutipes.

• Mycobiology
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• 제41권1호
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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.

• 한국균학회지
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• 제39권3호
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• pp.235-238
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• 2011

팽이균사체의 형질전환을 위하여 Agrobacterium 세포를 사용하였다. 특히, Agrobacterium 세포의 감염단계 전에 약한 NaOH용액을 처리하였으며 이로써 균사체 세포들의 표면 상해 발생을 기대하였다. 그 결과, hygromycin 저항성 ($hyg^r$) 균사체는 NaOH 처리를 거친 경우에서만 출현하였다. 형질전환 균사체의 $hyg^r$ 유전자 도입은 PCR로 확인되었으며 또한 Southern blot hybridization과 western blotting 분석에 의하여 단일 유전자 copy의 삽입과 외래유전자의 발현을 확인할 수 있었다. 본 연구는 팽이균사체에 대한 효율적인 Agrobacterium 이용 형질전환수단을 보여주고 있다.

• Mycobiology
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• 제42권4호
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• pp.322-330
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• 2014

The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges, fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylation sites (Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1 (Asn-454), fvLac-2 (Asn-437 and Asn-455), fvLac-3 (Asn-111 and Asn-237), and fvLac4 (Asn-402 and Asn-457). In addition, the first 19~20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction analyses clearly reveal that $CuSO_4$ affects the induction and the transcription level of these laccase genes.