• Microbiology and Biotechnology Letters
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• v.47 no.2
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• pp.250-258
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• 2019

Peniphora sp. JS17, isolated from forest old tree, produced extracellular enzymes that decolorized human hair melanin. The JS17 strain had laccase and manganese peroxidase activity while it did not has lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS17 strain originated from laccase. The culture conditions to maximize the production of melanin bleaching enzymes from Peniophora sp. JS17 mycelia were investigated. Among the tested media for the laccase production, minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$) showed the highest activity of laccase. Then, to optimize the culture condition for the laccase activity, the influence of various carbon and nitrogen sources was investigated in minimal medium. Among various carbon and nitrogen sources, 2% xylose and 0.4% tryptone showed the highest production of laccase, respectively. The enzyme was purified using $(NH_4)_2SO_4$ precipitation and Hitrap Q sepharose column, and the purified enzyme showed two isoenzymatic bands with molecular masses of about 70 kDa by SDS-PAGE. The melanin decolorization activity was 77% and 55% within 48 h in the presence of 1-hydroxybenzotriazole (HBT) and syringaldehyde, respectively, whereas only about 9% melanin decolorized in case of no mediator.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.463-470
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• 2020

The JS18 strain was isolated from an old tree forest and produced extracellular enzymes that decolorize synthetic melanin. Phylogenetic analysis, based on the internal transcribed spacer (ITS) sequence, indicate that JS18 belongs to the Trametes velutina species. JS18 demonstrated laccase activity but no manganese peroxidase or lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS18 strain originated from the laccase. Syringic acid and CuSO4 induced maximum laccase production, yielding 98 U/ml laccase activity after cultivation for 7 days at 25℃. T. velutina secretes an extracellular laccase in GYP medium, and this enzyme was purified using (NH4)2SO4 precipitation, Hi-trap Q Sepharose columns and gel filtration. The molecular weight of the purified enzyme was estimated to be 67 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme produced 80% of its melanin decolorization activity within the first 24 h of evaluation in the presence of 1-hydroxybenzotriazole (HBT), while only about 4% of the melanin was decolorized in the absence of the mediator. The greatest decolorization was observed at 1.5 mM/l HBT, which decolorized 81% of the melanin within the first 24 h. The optimum pH and temperature for this decolorization were found to be 5.0 and 37℃, respectively. Our results suggest the possibility of applying HBT induced T. velutina JS18 laccase-catalyzed melanin decolorization.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.93-95
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• 2010

White rot fungi which have lignin degrading enzymes show high degrading activity to diverse recalcitrant compounds such as polycyclic aromatic compounds, dyes, explosives and endocrine disrupting chemicals. We have examined decolorizing activity of dyes by Phlebia tremellosa and two transformants which had genetically transformed using laccase or manganese peroxidase (MnP) gene. In case of methyl green, wild type strain showed 50% decolorization while laccase transformant (TF2-1) and MnP transformant (T5) showed more than 90% decolorization on day 3. Remazol brilliant blue R(RBBR) was decolorized up to 85% by two transformants while the wild type showed 67% decolorization on day 3. Transformants TF2-1 and T5 both showed increased laccase and MnP activity respectively during the whole growing phase.

• 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.

• Journal of Mushroom
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• v.20 no.1
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• pp.29-33
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• 2022

In this study, five different Lentinula edodes cultivar (Chamaram, Sanbaekhyang, Sanjo 713ho, Sanjo 715ho, Sanjo 718ho) were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in MEB medium, respectively. Chamaram and Sanjo 713ho decolorized RBBR rapidly in MEB medium within 3 and 5 days. The activities of manganese peroxidase (MnP) and laccase were determined on the MEB medium with and without lignin. Sanjo 713ho resulted the highest ligninolytic enzyme activities on incubation day 1, indicating of 1,213 U/mg of MnP activity and 1,421 U/mg of laccase activity.

• Journal of The Korean Society of Grassland and Forage Science
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• v.34 no.1
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• pp.15-20
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• 2014

To investigate lignification process and its physiological significance under water-deficit condition, the responses of peroxidases, polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) in relation to leaf water status to the short term of water deficit treatment in the leaves with different maturities in forage rape were measured. The significant decrease in relative water content (RWC) and leaf osmotic potential (${\Psi}{\pi}$) were apparent after 5 d of water-deficit treatment. The activity of guaiacol peroxidase (GPOD), ascorbate peroxidase (APOD), coniferyl alcohol peroxidase (CPOD), and syringaldazine peroxidase (SPOD) was depressed especially in middle and old leaves when compared with that of control leaves. On the other hand, in young leaves, a significant increase in CPOD (+34%) and SPOD (+24%) activity as affected by water-deficit treatment was apparent. The activation of PAL and PPO was observed in middle and old leaves for PAL and in young and middle leaves for PPO. These results suggest that peroxidases in middle and old leaves did not involve in lignification under mild water-deficit stress, whereas CPOD and SPOD in young leaves participate in lignification by a coordination with PAL and PPO to incorporate phenol and lignin into the cell walls.

• Korean Journal of Microbiology
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• v.55 no.2
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• pp.177-179
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• 2019

Pseudomonas kribbensis CHA-19 was isolated from a temperate forest soil (mid latitude) in New Jersey, USA, for its ability to degrade humic acids, a main component of humic substances (HS), and subsequently confirmed to be able to decolorize lignin (a surrogate for HS) and catabolize lignin-derived ferulic and vanillic acids. The draft genome sequence of CHA-19 was analyzed to discover the putative genes for depolymerization of polymeric HS (e.g., dye-decolorizing peroxidases and laccase-like multicopper oxidases) and catabolic degradation of HS-derived small aromatics (e.g., vanillate O-demethylase and biphenyl 2,3-dioxygenase). The genes for degradative activity were used to propose a HS degradation pathway of soil bacteria.

• 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 Microbiology and Biotechnology
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• v.10 no.3
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• pp.344-348
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• 2000

Abstract The removal percentage (94%) of 100 ppm of pyrene in a shaken culture of white rot fungus, Irpex lacteus, was much higher than that in a static culture (37.9%). Over 90% of the pyrene disappeared with I. lacteus grown at $15-27^{\circ}C$, yet less than 50% was removed at $37^{\circ}C$. The transformation rates of pyrene ($4.5-5.0{\;}\mu\textrm{g}/ml/day$) were not very different among cultures with 5- 30% inoculum sizes, and over 90% of the 100 ppm pyrene was removed in every case during 20 days of incubation. The biodegradation of pyrene by I. lacteus was confirmed by measuring the $CO_2$ evolved from the mineralization of the added pyrene. The activity of lignin peroxidase (LiP), which is known to be involved in the biodegradation by white rot fungi, was high between 8 to 12 days of incubation. Although manganese peroxidase activity was demonstrated during the same period as LiP, its activity was quite low, and no laccase activity was detected. Even though the activity patterns of ligninolytic enzymes did not coincide with the pyrene removal, this study shows that I. lacteus has a high biodegrading capability and can be a candidate for the bioremediation of polycyclic aromatic hydrocarbon contaminants.inants.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.83-85
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• 2019

Pseudomonas sp. PAMC 29040 was isolated from a maritime tundra soil in Antarctica for its ability to degrade lignin and subsequently confirmed to be able to depolymerize heterogeneous humic substance (HS), a main component of soil organic matter. The draft genome sequences of PAMC 29040 were analyzed to discover the putative genes for depolymerization of polymeric HS (e.g., dye-decolorizing peroxidase) and catabolic degradation of HS-derived small aromatics (e.g., vanillate O-demethylase). The information on degradative genes will be used to finally propose the HS degradation pathway(s) of soil bacteria inhabiting cold environments.