• The Korean Journal of Mycology
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• v.25 no.3 s.82
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• pp.191-201
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• 1997

A mutant (HSMl) of Cryphonectria parasitica created during transformation reproduced the hypovirulent symptoms in virus-free wild type. Its phenomena have been proved with morphological marker such as reduced sporulation, pigmentation, and laccase production. In addition to the changes in phenotypic characteristics, down-regulations of Lac1, Crp1, Vir1 and Vir2 were also observed. The integration of transforming vector was confirmed and located within genome by marker rescuing. Vector integration occurred between two genes, Cpg2 and Cpg3, which resulted in the disruption of neither Cpg2 nor Cpg3. Both Cpg2 and Cpg3 genes, sized at 1.8 kb and 1.9 kb respectively, were rarely transcribed genes in Cryphonectria parasitica. Cpg2 expression was significantly overexpressed from 4 to 5 day old culture of both UEP1 and HSM1 while no differences were observed in Cpg3 expression. It appears that an aberration from the normal expression of Cpg2, not Cpg3, results in the hypovirulent symptoms in virus-free wild type.

• The Korean Journal of Mycology
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• v.43 no.1
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• pp.26-32
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• 2015

In this study, a new cultivar of Lentinula edodes was bred from dikaryotic KFRI 490 and monokaryotic KFRI 536 by Di-mon method. The new breed was named "Poongnyunko" and physiological characteristics was investigated. Its optimal growing temperature was $25^{\circ}C$, and it had thin density. Its colony was colorless and did not form any epithelium. On the $7^{th}$ day of mycelial growth, "Poongnyunko" was 56.6 mm, and showed higher cellulase activity compared with "Sanlim 4-ho"(52.9 mm). On the $5^{th}$ day of mycelial growth, "Poongnyunko" was 58.0 mm, and showed higher laccase activity compared with "Sanlim 4-ho" (55.6 mm). Its pH at a substrate was 6.0. Before cultivation, the pH of sawdust substrate was 4.7-5.5 went down to 3.7-3.9 after 120-day cultivation. After 60 days of cultivation, the change of $CO_2$ was the greatest. During the cultivation, $CO_2$ concentration in a growing bag was 4.67% to 3.90%.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.507-513
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• 2017

Bacillus subtilis spores can be used for protein display to engineer protein properties. This method overcomes viability and protein-folding concerns associated with traditional protein display methods. Spores remain viable under extreme conditions and the genotype/phenotype connection remains intact. In addition, the natural sporulation process eliminates protein-folding concerns that are coupled to the target protein traveling through cell membranes. Furthermore, ATP-dependent chaperones are present to assist in protein folding. CotA was optimized as a whole-cell biocatalyst immobilized in an inert matrix of the spore. In general, proteins that are immobilized have advantages in biocatalysis. For example, the protein can be easily removed from the reaction and it is more stable. The aim is to improve the pH stability using spore display. The maximum activity of CotA is between pH 4 and 5 for the substrate ABTS (ABTS = diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). However, the activity dramatically decreases at pH 4. The activity is not significantly altered at pH 5. A library of approximately 3,000 clones was screened. A E498G variant was identified to have a half-life of inactivation ($t_{1/2}$) at pH 4 that was 24.8 times greater compared with wt-CotA. In a previous investigation, a CotA library was screened for organic solvent resistance and a T480A mutant was found. Consequently, T480A/E498G-CotA was constructed and the $t_{1/2}$ was 62.1 times greater than wt-CotA. Finally, E498G-CotA and T480A/E498G-CotA yielded 3.7- and 5.3-fold more product than did wt-CotA after recycling the biocatalyst seven times over 42 h.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.347-357
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• 2019

This study evaluated the raw materials for spawning Perenniporia fraxinea, to eliminate urushiol. The growth rates of spawns on grains of millet, brown rice, and wheat were 4.92±0.05, 2.20±0.03, and 1.93±0.03 mm/day, respectively, and the laccase activity was 0.86±0.02, 0.04±0.01, and 0.01±0.00 U/mL, respectively. These observations revealed millet as the most appropriate grain for spawn production in terms of growth rate and enzyme activity. Inoculation of lacquer tree (Rhus verniciflua Stokes) stem bark with millet spawns of P. fraxinea resulted in a reduction of its urushiol contents, up to 86.6% on the third day and up to 98.5% on the seventh day. The optimal period of cultivation to eliminate urushiol was three days with 68% of residual flavonoids and 42% of phenolic components. When compared to the product cultivated from liquid spawn, the millet spawn reduced the cultivation period from 10 days to 3 days for eliminating urushiol.

• Mycobiology
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• v.29 no.3
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• pp.173-175
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• 2001

The fruit body of Ramaria botrytis DGUM 29001 was used to determine enzyme activities of fruit body. The specific activity of laccase was the highest(6.5 unit/mg$\cdot$protein) and that of $\alpha$-amylase and xylanase was relatively high. However, little or no enzyme activity of $\beta$-glucosidase, CMCase, exo-$\beta$-1,4-glucanase, chitinase, lipase and protease was found.

• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.823-832
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• 2021

Mushroom cultivation along with the palm oil industry in Malaysia have contributed to large volumes of accumulated lignocellulosic residues that cause serious environmental pollution when these agroresidues are burned. In this study, we illustrated the utilization of lignocellulolytic enzymes from the spent mushroom substrate of Pleurotus pulmonarius for the hydrolysis of palm oil mill effluent (POME). The hydrolysate was used for the production of biohydrogen gas and enzyme assays were carried out to determine the productivities/activities of lignin peroxidase, laccase, xylanase, endoglucanase and β-glucosidase in spent mushroom substrate. Further, the enzyme cocktails were concentrated for the hydrolysis of POME. Central composite design of response surface methodology was performed to examine the effects of enzyme loading, incubation time and pH on the reducing sugar yield. Productivities of the enzymes for xylanase, laccase, endoglucanase, lignin peroxidase and β-glucosidase were 2.3, 4.1, 14.6, 214.1, and 915.4 U g^-1, respectively. A maximum of 3.75 g/lof reducing sugar was obtained under optimized conditions of 15 h incubation time with 10% enzyme loading (v/v) at a pH of 4.8, which was consistent with the predicted reducing sugar concentration (3.76 g/l). The biohydrogen cumulative volume (302.78 ml H₂.L^-1 POME) and 83.52% biohydrogen gas were recorded using batch fermentation which indicated that the enzymes of spent mushroom substrate can be utilized for hydrolysis of POME.

• KSBB Journal
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• v.18 no.5
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• pp.398-403
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• 2003

Due to the low biodegradability of dyes, conventional biological wastewater treatment systems are inefficient in treating dye wastewater. Various white-rot fungi were investigated for the decolorization of six industrial dyes (reactive blue 5, reactive blue 16, reactive black 5, acid black 52, reactive orange 16, and acid violet 43). Among ten fungi, T. versicolor KCTC 16781 was selected as a testing strain because this had the best performance of decolorization for six dyes from the results of the solid culture experiments. In liquid culture medium, T. versicolor KCTC 16781 decolorized over 96% of six dyes for 48 hrs. Laccase started to produce in the early stage of the culture, and showed the highest peak value of 2.3 U/mL in 24 hrs. Enzyme activity remained constant until the end of culture. Fungal decolorization is a promising alternative to replace or supplement present treatment process.

• Food Science and Biotechnology
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• v.16 no.6
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• pp.935-942
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• 2007

The stem bark of Rhus verniciflua (RVSB) has been used in herbal medicine to treat diabetes mellitus and stomach ailments for thousands of years in Korea, despite its content of the plant allergen, urushiol. A new biological approach for the removal of urushiol from RVSB using mushrooms is described. All mushroom species (11 sp.) employed in this study were able to grow on RVSB, although the growth rate (mm/day) was lower than the control (sawdust). The components of urushiol congeners [C15 triene (m/z 314), C15 diene (m/z 316), C15 monoene (m/z 318), and C15 saturated (m/z 320)] were purified by HPLC and identified by GC-MS. A C15:3 (3-pentadecatrienly catechol) was found to be most abundant in RVSB. Urushiol analogues decreased remarkably from 154.15 to 10.73 mg/100 g (approximately 93%) by Fomitella fraxinea, whereas Trametes vercicolor showed only a 1.46% degradation capacity despite its 2 fold higher growth rate. Similarly, laccase activity was found to be high for F. fraxinea and low for T. vercicolor. Moreover, approximately 98% detoxification was accomplished by F. fraxinea cultivated on RVSB supplemented with 20%(w/w) rice bran. These findings suggest that mushrooms can be used in the detoxification of RVSB.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.107-109
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• 2008

Medicinal fungi, Phellinus linteus and Inonotus xeranticus, produce a cluster of yellow pigment in their fermentation broth that acts as an important element of biological activity. The pigment is composed of diverse polyphenols with a styrylpyrone moiety, mainly hispidin and its dimers, 3,14'-bihispidinyl, hypholomine B, and 1,1-distyrylpyrylethan. Although dimeric hispidins were proposed to be biosynthesized from two molecules of monomer via oxidative coupling by ligninolytic enzymes, laccase and peroxidase, the details of this process remain unknown. In this preliminary study, we attempted to achieve enzymatic synthesis of the hispidin dimer from hispidin by using commercially available horseradish peroxidase (HRP). Consequently, a hispidin dimer, 3,14'-bihispidinyl, was synthesized, whereas the other dimers, hypholomine B and 1,1-distyrylpyrylethan, were not produced. This result suggested that the oxidative coupling at the C-3 and C-14' positions of hispidins was dominant in the process of dimerization by HRP, and indicated that additional catalysts or substrates would be needed to synthesize other hispidin dimers present in the fungal metabolite.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.159-161
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• 2005

Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.