• Title/Summary/Keyword: manganese-dependent peroxidase (MnP)

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Characterization of Laccase Purified from Korean Pycnoporus cinnabarinus SCH-3 (한국산 주걱송편버섯(Pycnoporus cinnabarinus) SCH-3로부터 정제 된 Laccase의 특성)

  • Park, Eun-Hye;Yoon, Kyung-Ha
    • 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.

Screening of Biodegradable Function of Indigenous Ligno-degrading Mushroom Using Dyes

  • Jang, Kab-Yeul;Cho, Soo-Muk;Seok, Soon-Ja;Kong, Won-Sik;Kim, Gyu-Hyun;Sung, Jae-Mo
    • 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).

Biodegradation of 4-t-Octylphenol by Basidioradulum molare and Schizopora paradoxa and Estrogenecity Reduction of its Metabolites (옥틸페놀(4-t-Octylphenol)의 Basidioradulum molare와 Schizopora paradoxa에 의한 분해 및 에스트로겐성 저감효과)

  • Lee, Soo-Min;Ku, Bon-Wook;Lee, Jae-Won;Choi, Don-Ha;Jeung, Eui-Bae;Choi, In-Gyu
    • 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.