• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.347-358
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• 2011

Fomitopsis palustris, a brown-rot basidiomycete, causes the most destructive type of decay in wooden structures. In spite of its great economic importance, very little information is available at the molecular level regarding its complex decay process. To address this, we generated over 3,000 expressed sequence tags (ESTs) from a cDNA library constructed from F. palustris. Clustering of 3,095 high-quality ESTs resulted in a set of 1,403 putative unigenes comprising 485 contigs and 918 singlets. Homology searches based on BlastX analysis revealed that 78% of the F. palustris unigenes had a significant match to proteins deposited in the nonredundant databases. A subset of F. palustris unigenes showed similarity to the carbohydrateactive enzymes (CAZymes), including a range of glycosyl hydrolase (GH) family proteins. Some of these CAZyme-encoded genes were previously undescribed for F. palustris but predicted to have potential roles in biodegradation of wood. Among them, we identified and characterized a gene (FpCel45A) encoding the GH family 45 endoglucanase. Moreover, we also provided functional classification of 473 (34%) of F. palustris unigenes using the Gene Ontology hierarchy. The annotated EST data sets and related analysis may be useful in providing an initial insight into the genetic background of F. palustris.

• Microbiology and Biotechnology Letters
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• v.46 no.3
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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.

• YAKHAK HOEJI
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• v.38 no.2
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• pp.158-165
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• 1994

A protein-polysaccaride fraction Kp(53.9% polysaccharide, 14.2% protein) was separated from the shake-cultured mycelia of a basidiomycetous fungus, Phellinus linteus, and its antitumor activity against sarcoma 180 in ICR mice was investigated. When administered after the tumor implantation, Kp exerted antitumor activity by inhibiting the growth of the sarcoma 180 solid tumor by 71.5% and increasing the life span of the sarcoma 180 ascitic mice by 51.5% at 100 mg/kg. In pretreatment tests, in which Kp was administered once daily for 9 days before the tumor implantation, Kp inhibited the growth of the solid and ascites form of sarcoma 180, respectively, by 35.4% and by 80.3% at 100 mg/kg. However, Kp showed no in vitro cytotoxic activity against a murine leukemia L1210 and a human gastric tumor SNU.1 upto the concentration of $200\;{\mu}g/ml$. From these results, it is clear that the antitumor activity of Kp is exerted through its immunomodulating activity on the host's immune system.

• The Korean Journal of Mycology
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• v.17 no.1
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• pp.35-38
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• 1989

Gene libraries of DNA from Flammulina velutipes were constructed using Escherichia coli plasmid pBR 322. Leu 2 gene coding ${\beta}-isopropylmalate$ dehydrogenase from F. velutipes was cloned by complementation of leucine requiring mutant of E. coli. The size of inserted DNA fragment of this clone is about 1 Kbp. The fragment has Bam H1 and Ava 1 restriction sites.

• Journal of the Korean Wood Science and Technology
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• v.27 no.4
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• pp.1-14
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• 1999

As the biodegradation of wood constituents has been understood as a multi-basidiomycetes and enzymatic processes, this review will focus on the roles of low molecular compounds and radicals working in harmony with fungal enzymes. Wood rotting basidiomycete fungi penetrate wood, and lead to more easily metabolize carbohydrates of the wood complex. The white-rot fungi, having versatile enzymes, are able to attack directly the "lignin barrier". They also use a multi-enzyme system including so-called "feedback" type enzymes allowing for simultaneous degradation of lignin and carbohydrates. The multi-enzymes including laccase support the proposed route by explaining how the high molecular weight enzymes can function in the wood complex. These enzymes may function separately or cooperate each other. In addition, veratryl alcohol oxidase, cellobiose dehydrogenase, arylalcohol dehydrogenase, and particularly low molecular mediators and radicals have an important role in wood biodegradation. However, the possibility of other mechanism as well as other enzymes, as operating as feedback systems in the process of wood degradation, could not be excluded.

• Mycobiology
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• v.37 no.3
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• pp.161-170
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• 2009

Cryptococcus neoformans is a basidiomycete human fungal pathogen that causes meningoencephalitis in both immunocompromised and immunocompetent individuals. The ability to sense and respond to diverse extracellular signals is essential for the pathogen to infect and cause disease in the host. Four major stress-activated signaling (SAS) pathways have been characterized in C. neoformans, including the HOG (high osmolarity glycerol response), PKC/Mpk1 MAPK (mitogen-activated protein kinase), calcium-dependent calcineurin, and RAS signaling pathways. The HOG pathway in C. neoformans not only controls responses to diverse environmental stresses, including osmotic shock, UV irradiation, oxidative stress, heavy metal stress, antifungal drugs, toxic metabolites, and high temperature, but also regulates ergosterol biosynthesis. The PKC(protein kinase C)/Mpk1 pathway in C. neoformans is involved in a variety of stress responses, including osmotic, oxidative, and nitrosative stresses and breaches of cell wall integrity. The $Ca^{2+}$/calmodulin- and Ras-signaling pathways also play critical roles in adaptation to certain environmental stresses, such as high temperature and sexual differentiation. Perturbation of the SAS pathways not only impairs the ability of C. neoformans to resist a variety of environmental stresses during host infection, but also affects production of virulence factors, such as capsule and melanin. A drug(s) capable of targeting signaling components of the SAS pathway will be effective for treatment of cryptococcosis.

• Archives of Pharmacal Research
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• v.10 no.3
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• pp.179-183
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• 1987

To investigate antitumor constituents of higher fungi, the carpophores of Polyporus giganteus Pers. ex. Fr. (81 g, dry weight) which were collected in Indiana, U. S. A. were examined for antitumor activity. Two protein-bound-polysaccharide fractions (I and II) were prepared from the hot water extract and one fraction (III) from the 0.1 N NaOH extract of the carpophores. The antitumor effect of each fraction was tested against sarcoma 180 implanted subcutaneously in female ICR mice. Of three fractions, Fraction II showed 85.2% inhibition ratio at the dose of 20 mg/kg/day for 10 days and was named gigantan. Gigantan was found to contain 59% polysaccharide and 27% protein. Its polysaccharide moity was a heteroglycan that consisted of mainly glucose (89.3%), galactose (7.7%), minaose (2.0%) and fructose (1.0%).

• Archives of Pharmacal Research
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• v.20 no.3
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• pp.206-211
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• 1997

The optimal conditions for the production and regeneration of the protoplasts from Lentinula edodes were studied. Protoplast formation from the mycelia of L. edodes which were cultured in liquid medium showed a significantly high yield compared with that of the mycelia which were cultured on cellophane covered agar media. A mixture of Novozyme 234 (15 mg/ml) and Cellulase Onozuka R10 (10 mg/ml) in 0.6 M mannitol (pH 4) was optimal lytic enzyme for the protoplast release. The optimal incubation time and mycelia age were 3.5-4 hours at $30^{\circ}C$ and 6-8 days, respectively. Regeneration frequency was 0.18% plated onto a medium containing 0.6 M sucrose, and 0.08% plated onto a medium containing mannitol. But hardly any regeneration was observed in the media containing NaCl, KCl, or $MgSO_{4}$ More than 90% of the protoplasts contained nuclei and the nucleus number per protoplast was 1.1. The DNA content per nucleus was 5.1 pg. The diameter of the protoplast was $3-5{\mu}m$ and it had a well defined cell structure.

• Archives of Pharmacal Research
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• v.19 no.3
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• pp.207-212
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• 1996

To find pharmacologically active components of Agrocybe cylin(Iracea, its basidiocarps were extracted with water. The extracts were separated by DEAE cellulose column chromatography, Sepharose CL-4B gel filtration, and Concanavalin A-Sepharose 4B affinity chromatography. Among the obtained fractions from A, cylinclracea, fraction IN which was the neutral proteinbound-polysaccharide fraction exhibited a marked antitumor activity and it was tentatively named "Cylindan". It showed about 70% of tumor inhibition against the solid form of sarcoma 180 when a dose of 30 mg/kg/day was intraperitoneally injected into ICR mice. When each fraction was examined by chemical analysis, Cylindan consisted of 85% polysaccharide, 3% protein and 1% hexosamine. Its polysaccharide moiety contained glucose, mannose, fucose and galactose and its protein moiety contained the comparatively large amounts of aspartic acid and glycine, and other 11 amino acids.

• Journal of Microbiology
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• v.39 no.2
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• pp.121-125
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• 2001

Methanol extract prepared from the fruitbody of Phellinus linteus (EPL) showed anti-tumor and immune-stimulating activities. The invasion activity of Bl6-F10 melanoma cells through a reconstituted basement membrane to the collagen-coated lower surface of the filters was inhibited about 67% by EPL (100 $\mu\textrm{g}$/ml). Also, EPL inhibited the expression of the mRNA for MMP-2 and MMP-9. In vivo treatment of C57BL/6 mice (150 mg/kg) with EPL for 14 days, the pulmonary colonization was found to be inhibited about 75%. Using reverse transcriptionpolymerase chain reaction (RT-PCR) analysis, we found that cytokine IL-12 and INF-${\gamma}$ genes were induced by EPL. Furthermore, EPL stimulated the proliferation of CD4$\^$+/(33.5%) and CD8$\^$+/(17.7%) in mouse splenocytes.