• Journal of the Korean Wood Science and Technology
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• v.26 no.3
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• pp.63-72
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• 1998

In order to evaluate the biodegradability and mechanism of 4,5,6-trichloroguaiacol (TCG) produced from bleaching process in pulp mill by Phanerochaete chrysosporium, Trametes versicolor, and Inonotus cuticularis, changes in TCG and its metabolites during biodegradation were analyzed by HPLC, and GC/MS spectrometry. By three fungi, the maximum biodegradability against TCG were very quickly reached, compared with other chlorinated aromatic compounds such as PCP. Within 24 hrs, T versicolor indicated up to 95% of TCG removal rate, and P. chrysosporium and I. cuticularis also showed more than 80%, and 90%, respectively. Particularly, in case of T. versicolor, the removal rate of TCG after 1 hr. incubation was reached to approximately 90%, implying very rapid metabolization of TCG. However, by analyzing the filtrates extracted from TCG containing culture by GC/MS, the major metabolites at initial stage of biodegradation were dimers, indicating that the added TCG monomers were quickly polymerized. The others were trichloroveratrole, dichloroguaiacol, and trichlorobenzoic acid, suggesting that TCG may be biodegraded by several sequential reactions such as polymerization, oxidation, methylation, dechlorination, and hydroxylation. In other experiments, the extracellular fluid which did not contain any fungal mycelia was used to evaluate the effect of mycelia on TCG biodegradation. The extracellular fluid of T. versicolor also biodegraded TCG up to 90% within 24hrs, but those of P. chrysosporium and I. cuticularis did not show any good biodegradability. T versicolor showed the highest value of laccase, and other two fungi indicated a little activity of lignin peroxidase (LiP) and manganese peroxidase (MnP). In addition, the laccase activity of T. versicolor was very linearly proportional to the removal rate of TCG during incubation, in other words, showing the induction effect against TCG. Consequently, the biodegradation of TCG was very dependent upon the activity of laccase.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.59-66
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• 2003

Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

• Korean Journal of Environmental Biology
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• v.34 no.3
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• pp.201-207
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• 2016

The hazards associated with the polycyclic aromatic hydrocarbons (PAHs) are known to be recalcitrant by their structure, but white rot fungi are capable of degrading recalcitrant organic compounds. Phlebia brevispora KUC9045 isolated from Korea was investigated its efficiency of degradation of four PAHs, such as phenanthrene, anthracne, fluoranthene, and pyrene. And the species secreted extracellular laccase and MnP (Manganese dependent peroxidase) during degradation. P. brevispora KUC9045 demonstrated effective degradation rates of phenanthrene (66.3%), anthracene (67.4%), fluoranthene (61.6%), and pyrene (63.3%), respectively. For enhancement of degradation rates of PAHs by the species, Remazol Brilliant Blue R (RBBR) was preferentially supplemented to induce ligninolytic enzymes. The biodegradation rates of the three PAHs including phenanthrene, fluoranthene, and pyrene were improved as higher concentration of Remazol Brilliant Blue R was supplemented. However, anthracene was degraded with the highest rate among four PAHs after two weeks of the incubation without RBBR addition. According to the previous study, RBBR can be clearly decolorized by P. brevispora KUC9045. Hence, the present study demonstrates simultaneous degradation of dye and PAHs by the white rot fungus. And it is considered that the ligninolytic enzymes are closely related with the degradation. In addition, it indicated that dye waste water might be used to induce ligninolytic enzymes for effective degradation of PAHs.

• Mycobiology
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• v.43 no.3
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• pp.297-302
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• 2015

Two white rot fungi, Ceriporia sp. ZLY-2010 (CER) and Stereum hirsutum (STH) were used as biocatalysts for the biotransformation of (-)-${\alpha}$-pinene. After 96 hr, CER converted the bicyclic monoterpene hydrocarbon (-)-${\alpha}$-pinene into ${\alpha}$-terpineol (yield, 0.05 g/L), a monocyclic monoterpene alcohol, in addition to, other minor products. Using STH, verbenone was identified as the major biotransformed product, and minor products were myrtenol, camphor, and isopinocarveol. We did not observe any inhibitory effects of substrate or transformed products on mycelial growth of the fungi. The activities of fungal manganese-dependent peroxidase and laccase were monitored for 15 days to determine the enzymatic pathways related to the biotransformation of (-)-${\alpha}$-pinene. We concluded that a complex of enzymes, including intra- and extracellular enzymes, were involved in terpenoid biotransformation by white rot fungi.

• Journal of Microbiology
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• v.44 no.2
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• pp.177-184
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• 2006

The characteristic biodegradation of monomeric styrene by Phanerochaete chrysosporium KFRI 20742, Trametes versicolor KFRI 20251 and Daldinia concentrica KFRI 40-1 was carried out to examine the resistance, its degradation efficiency and metabolites analysis. The estrogenic reduction effect of styrene by the fungi was also evaluated. The mycelium growth of fungi differentiated depending on the concentration levels of styrene. Additionally P. chrysosporium KFRI 20742 showed superior mycelium growth at less than 200 mg/l, while D. concentrica KFRI 40-1 was more than 200 mg/l. The degradation efficiency reached 99 % during one day of incubation for all the fungi. Both manganese-dependent peroxidase and laccase activities in liquid medium were the highest at the initial stage of incubation, whereas the lowest was after the addition of styrene. However, both activities were gradually recovered after. The major metabolites of styrene by P. chrysosporium KFRI 20742 were 2-phenyl ethanol, benzoic acid, cyclohexadiene-1,4-dione, butanol and succinic acid. From one to seven days of incubating the fungi, the expression of pS2 mRNA widely known as an estrogen response gene was decreased down to the level of baseline after one day. Also, the estrogenic effect of styrene completely disappeared after treatment with supernatant of P. chrysosporium KFRI 20742 from one week of culture down to the levels of vehicle.

• The Korean Journal of Mycology
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• v.32 no.2
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• pp.112-118
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• 2004

Laccase produced by Trametes hirsuta S1 isolated from Korea was partially purified and characterized using ultrafiltration, anion exchange chromatography and affinity chromatography. The laccase was produced as the predominant extracellular enzyme during primary metabolism. Neither lignin peroxidase nor veratryl alcohol oxidase (VAO) were detected in the culture fluid. Addition of 2,5-xylidine enhanced 4-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 12%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to $51.2\;{\mu}M\;and\;56.8\;{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $50^{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 20 min at $70^{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the T. hirsuta S1 laccase showed 100% of homology to those of laccase from C. hirsutus.

• Mycobiology
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• v.37 no.1
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• pp.53-61
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• 2009

The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

• Journal of the Korean Wood Science and Technology
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• v.32 no.4
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• pp.27-35
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• 2004

Recalcitrant 4-t-Octylphenol used as a surfactant was subjected to the biodegradation with wood rot fungi, Basidioradulum molare and Schizopora paradoxa. Two fungi were grown in the culture medium containing various concentrations of 4-t-Octylphenol in order to investigate their resistance against 4-t-octylphenol Schizopora paradoxa was reached to the full growth within 14 incubation days in the concentration of more than 200 ppm of 4-t-Octylphenol, while Basidioradulum molare showed the inhibitory mycelium growth as the concentration was increased Schizopora paradoxa and Basidioradulum molare biodegraded 95% and 36% of initial concentration of 4-t-Octylphenol at first incubation day, respectively. However, the biodegradation capability reached to more than 95% after 3 incubation days. During the biodegradation of 4-t-Octylphenol, the activity of manganese dependent peroxidase was induced by the addition of 4-t-Octylphenol in the culture medium of Schizopora paradoxa, but that of laccase was maximal before the addition. The reduction of estrogenecity was assayed by MCF-7 cell proliferation test and measurement of pS2 mRNA expression. The level of pS2 mRNA was decreased down to the level of baseline at first incubation day. Also, estrogenecity of 4-t-Ocrylphenol completely disappeared after treatment with supernatant by Schizopora paradoxa and Basidioradulum molare from first incubation day of culture down to the levels of vehicle.