• Journal of Mushroom
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• v.19 no.4
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• pp.316-321
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• 2021

Ligninolytic enzymes were produced by Pleurotus ostreatus No.42, cultivated in a new kind of bioreactor that has a rotating draft tube with a helical ribbon. Maximum laccase (Lac) production (about 8,200 U/bioreactor) was reached after 3 days of incubation, then production decreased. Production of manganese peroxidase (MnP) in this fermenter reached a maximum level of about 8,400 U/bioreactor after 6 days of incubation. Lignin peroxidase (LiP) was not detected under these growth conditions. These results indicate that the rotary draft tube bioreactor (RTB) is compatible with large scale production of ligninolytic enzymes. MnP produced under these fermentation conditions was purified via a multistep process that included chromatography on Sepharose CL-6B, prep grade Superdex 75, and Mono-Q. This major isoenzyme was confirmed to have an apparent molecular weight of 36,400 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and its isoelectric point (IEF) was determined to be 3.95. N-terminal sequencing of the major isoenzyme from this fermentation was identical to that reported for an MnP3 isoenzyme isolated under different cultivation conditions, including stationary and shaking culture.

• Journal of Mushroom
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• v.11 no.2
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• pp.87-91
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• 2013

When No. 42 strain of Pleurotus ostreatus was cultivated at five different media, MnP and Lac but no LiP activity was detected throughout the culture period in the media. The production of MnP and Lac by No. 42 strain of Pleurotus ostreatus were correlated with wheat bran composition in the medium. In the liquid culture, maximum production of MnP and Lac were observed in the medium contained glucose-peptone- yeast-wheat bran(GPYW). However, in solid medium, maximum production of MnP was observed in wood meal-wheat bran(WMW) medium, but that of Lac was observed in wheat bran(W) medium.

• 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.

• 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.

• 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 Mushroom
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• v.7 no.3
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• pp.110-114
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• 2009

Sawdust bag cultivation of Shiitake mushroom (Lentinula edodes) is getting increase. The mycelia browning on the substrate surface is important for the stable production. The development of methods for the rapid mycelia browning is quite required. In this study changes in intracellular enzyme during the mycelial browning were investigated to find the rapid mycelia browning. Mycelia of L. edodes was changed into brown color while it grew in agar and liquid media like sawdust substrates. Mycelia of L. edodes was started to change color at 25 days after inoculation and browning was occurred in whole mycelia colony at 30 days and browning was completed at 40 days. The activities of enzymes was evaluated in these periodically color changing mycelia. Laccase activity was highest at 15 days after inoculation on PDB, but it gradually decreased from 15 days. Tyrosinase activity drastically increased in period between 30 days and 40 days while mycelia browning was progressed. The kinds of phosphotase identified by electrophoresis were esterase, acid phosphotase, and alkaline phosphotase. Activities of phosphotase were increased before the initiation of mycelial browning but they were decreased after browning.

• 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.

• The Korean Journal of Mycology
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• v.40 no.2
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• pp.98-103
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• 2012

This study has been conducted to screen the decolorization of 4 aromatic synthetic dyes and production of ligninolytic enzymes by 4 white rot fungi such as Bjerkanderia adusta, Cerrena unicolor, Pleurotus pulmonarius and Abortiporus biennis. It was found that B. adusta, C. unicolor, and P. pulmonarius have the ability to efficiently decolorize congo red and moderately decolorized amaranth and orange G in solid and liquid culture media. However, the decolorization rate of 4 synthetic dyes by A. biennis was relatively low. The decolorization of congo red, amaranth, orange G were related to the growth rate of the fungal mycelia in the solid medium. But, the all fungi tested did not efficiently decolorize methylene blue in the liquid culture media. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in 1% naphthalene supplemented potato dextrose broth medium. All fungi tested had the capability to produce laccase, lignin peroxidase and manganese peroxidase, and B. adusta was the best ligninolytic enzymes producing white rot fungus among other fungi tested.

• 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 Mushroom
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• v.14 no.2
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• pp.51-58
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• 2016

Basidiomycetous fungi are one of the most potent biodegraders because many of its species grow on dead wood or litter, in environments rich in lignocellulose. For the degradation of lignocellulose, basidiomycetes utilize their lignocellulytic enzymes, which typically include laccase (EC 1.10.3.2), lignin peroxidase (EC 1.11.1.14), xylanase (EC 3.2.1.8), and cellulase (EC 3.2.1.4). In recent years, the practical applications of basidiomycetes have ranged from the textile to the pulp and paper industries, and from food applications to bioremediation processes and industrial enzymatic saccharification of biomass. Recently, spent mushroom substrates of edible mushrooms have been used as sources of bulk enzymes to decolorize synthetic dyes in textile wastewater. In this review, the occurrence, mode of action, general properties, and production of lignocellulytic enzymes from mushroom species will be discussed. We will also discuss the potential applications of these enzymes.