• Title/Summary/Keyword: Phanerochaete chrysosporium

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Heterologous Expression of Phanerochaete chrysoporium Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green

  • Son, Yu-Lim;Kim, Hyoun-Young;Thiyagarajan, Saravanakumar;Xu, Jing Jing;Park, Seung-Moon
    • Mycobiology
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    • v.40 no.4
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    • pp.258-262
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    • 2012
  • cDNA of the glx1 gene encoding glyoxal oxidase (GLX) from Phanerochaete chrysosporium was isolated and expressed in Pichia pastoris. The recombinant GLX (rGLX) produces $H_2O_2$ over 7.0 nmol/min/mL using methyl glyoxal as a substrate. Use of rGLX as a generator of $H_2O_2$ improved the coupled reaction with recombinant manganese peroxidase resulting in decolorization of malachite green up to $150{\mu}M$ within 90 min.

Studies on the Ligninolytic Enzyme Activities During Biological Bleaching of Kraft Pulp with Newly Isolated Lignin-Degrading Fungi

  • Lee, Seon-Ho
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.2
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    • pp.8-14
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    • 1999
  • A screening has been performed to find hyper-ligninolytic fungi, which degtrade beech and pine lignin extensively in order to broaden the understanding of the ligninolytic enzymes elaborated by various white-rot fungi. One hundred and twenty two ligninolytic strains were selected from decayed woods with a selective medium for screening ligninolytic wood-rotting fungi. Two of them, Phanerochaete sordida YK-624 and YK-472, showed much higher ligninolytic activity and selectivity in beech-wood degradation than typical lignin-degrading fungi, phanerochaete chrysosporium and Coriolus versicolor. They also degraded birch dioxane lignin and residual lignin in unbleached kraft pulp(UKP) much more extensively than P. chrysosporium and C. versicolor. During fungal treatment of beech wood-powder, the fungus strain P. sordida YK-624 showed higher activity of extracellular manganese peroxidase (MnP) in the medium than P. chrysosporium. It also showed MnP activity, which would not be lignin peroxidast during treatment of oxygen-bleached kraft pulp(OKP) and under enzyme-inducing conditin.

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Bioconversion of Lignocellulose Materials

  • Pothiraj, C.;Kanmani, P.;Balaji, P.
    • Mycobiology
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    • v.34 no.4
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    • pp.159-165
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    • 2006
  • One of the most economically viable processes for the bioconversion of many lignocellulosic waste is represented by white rot fungi. Phanerochaete chrysosporium is one of the important commercially cultivated fungi which exhibit varying abilities to utilize different lignocellulosic as growth substrate. Examination of the lignocellulolytic enzyme profiles of the two organisms Phanerochaete chrysosporium and Rhizopus stolonifer show this diversity to be reflected in qualitative variation in the major enzymatic determinants (ie cellulase, xylanase, ligninase and etc) required for substrate bioconversion. For example P. chrysosporium which is cultivated on highly lignified substrates such as wood (or) sawdust, produces two extracellular enzymes which have associated with lignin deploymerization. (Mn peroxidase and lignin peroxidase). Conversely Rhizopus stolonifer which prefers high cellulose and low lignin containg substrates produce a family of cellulolytic enzymes including at least cellobiohydrolases and ${\beta}-glucosidases$, but very low level of recognized lignin degrading enzymes.

Expression of a Manganese Peroxidase Gene (mnp5) from White rot fungus Phanerochaete chrysosporium in the Pichia pastoris (백색부후균 Phanerochaete chrysosporium에서 유래한 Manganese Peroxidase Gene(mnp5)의 Pichia pastoris에서의 이종발현)

  • Lee, Jae-Won;Yang, In;Igarashi, Kiyohiko;Samejima, Masahiro;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.33 no.4 s.132
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    • pp.45-52
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    • 2005
  • The manganese peroxidase (mnp5) from white-rot fungus Phanerochaete chrysosporium has been heterologously expressed in the methylotrophic yeast Pichia pastoris. The majority of the rMnP5 (recombinant MnP5) produced by P. pastoris exhibited an approximate molecular mass 45 kDa considerably larger than that of the predicting mnp5 due to two glycosylation sites of mnp5. After site direct mutation treatment, the effect of N-linked hyperglycosylation was examined by enzyme activity. Analysis by sodium dodesyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining revealed a major protein band with a molecular mass of 37 kDa. Enzyme activity of M-rMnP5 (mutant recombinant MnP5) was similar to that of rMnP5, indicating that hyperglycosylation did not affect the active site. In this work, active mnp5 was successfully expressed in P. pastoris, suggesting that P. pastoris has potential capability of producing active heme-containing proteins.

Studies on the Recycling Technology of the Waste Paper with Wood Degradable Enzyme(I) - Separation of Crude Enzyme from Wood Degradable fungi - (목질분해효소에 의한 고지의 재활용 기술연구 (제1보) -목질분해균으로부터 조효소의 단리)

  • 양재경;이중명;엄태진
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.29 no.1
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    • pp.43-51
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    • 1997
  • The various culture conditions of Trichoderma viride(ATCC 3454) and Phanerochaete chrysosporium(ATCC 26921) with glucose-pepton medium, Mandels medium, YMG medium for wood degradable enzyme were examined. Mycellium of the two species grew profusely on glucose-pepton medium. Maximum fungal growth was observed about 10days. But CMCase, Fpase, laccase activity in the culture medium with glucose-pepton was not detected. When grown in fermenter culture using Mandels medium, Trichoderma viride produced CMCase and Fpase. Its CMCase activity was 0.15 lU/ml and Fpase activity was 0.3 IU/ml within about 4-6days. Phanerochaete chrysosporium grown in a YMG medium gave the best enzyme activity when they were grown under stationary culture with an atmosphere of 100% oxygen. Levels of laccase activity of 3.0 mull were achieved in stationary culture under 100% oxygen. The enzyme condensation by ultrafiltration method caused a 2-fold(cellulase) and 6-fold(laccase) as compared to control activity.

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Heterologous Expression of Endo-1,4-beta-xylanaseA from Phanerochaete chrysosporium in Pichia pastoris

  • Huy, Nguyen Duc;Thiyagarajan, Saravanakumar;Son, Yu-Lim;Park, Seung-Moon
    • Mycobiology
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    • v.39 no.2
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    • pp.121-124
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    • 2011
  • The cDNA of endo-1,$4-{\beta}-xylanaseA$, isolated from Phaenerocheate chrysosporium was expressed in Pichia pastoris. Using either the intrinsic leader peptide of XynA or the ${\alpha}$-factor signal peptide of Saccharomyces cerevisiae, xylanaseA is efficiently secreted into the medium at maximum concentrations of 1,946 U/L and 2,496 U/L, respectively.

Biodegradation of Pentachlorophenol by Various White Rot Fungi (수질분해균(水質分解菌)에 의한 Pentachlorophenol의 미생물분해(微生物分解))

  • Choi, In-Gyu;Ahn, Sye-Hee
    • Journal of the Korean Wood Science and Technology
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    • v.26 no.3
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    • pp.53-62
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    • 1998
  • In this research, 7 species of white rot fungi were used for determining the resistance against pentachlorophenol (PCP). Three fungi with good PCP resistance were selected for evaluating the biodegradability, and biodegradation mechanism by HPLC and GC/MS spectrometry. Among 7 fungi, there were significant differences on PCP resistance on 4 different PCP concentrations. In the concentrations of 50 and 100ppm ($\mu$g of PCP per g of 2% malt extract agar), most fungi were easily able to grow, and well suited to newly PCP-added condition, but in that of more than 250ppm, the mycelia growths of Ganoderma lucidum 20435, G. lucidum 20432, Pleurotus ostreatus, and Daldinia concentrica were significantly inhibited or even stopped by the addition of PCP to the culture. However, Trametes versicolor, Phanerochaete chrysosporium, and Inonotus cuticularis still kept growing at 250ppm, indicating the potential utilization of wood rot fungi to high concentrated PCP biodegradation. Particularly, P. chrysosporium even showed very rapid growth rate at more than 500ppm of PCP concentration. Three selected fungi based on the above results showed an excellent biodegradability against PCP. P. chrysosporium degraded PCP up to 84% on the first day of incubation, and during 7 days, most of added PCP were degraded. T. versicolor also showed more than 90% of biodegradability at 7th day, and even though the initial stage of degradation was very slow, I. cuticularis has been approached to 90% at 21 st day after incubation with dense growing pattern of mycelia. Therefore, the PCP biodegradability was definitely dependent on the rapid suitability of fungi to newly PCP-added condition. In addition, the PCP biodegradation by filtrates of P. chrysosporium, T. versicolor, and I. cuticularis was very minimal or limited, suggesting that the extracellular enzyme system may be not so significantly related to the PCP biodegradation. Among the biodegradation metabolites of PCP, the most abundant one was pentachloroanisole which resulted in a little weaker toxicity than PCP, and others were tetrachlorophenol, tetrachloro-hydroquinone, benzoic acid, and salicylic acid, suggesting that PCP may be biodegraded by several sequential reactions such as methylation, radical-induced oxidation, dechlorination, and hydroxylation.

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Kraft Pulping Characteristics by Bio-pretreatment with White-rot Fungus (백색부후균 생물 전처리에 의한 Kraft Pulp화 특성)

  • Kang, Kyu-Young;Jo, Byoung-Muk;Oh, Jung-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.4
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    • pp.103-110
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    • 2001
  • Bio-kraft pulping of Populus alba${\times}$glandulosa pretreated with white-rot fungus, Phanerochaete chrysosporium KCCM 34740, was investigated The biopulping efficiency was evalued based on fungal pretreatment time and properties of bio-kraft pulp (pulp yield, freeness, WRY, kappa number and brightness) in comparison to the controls. Pretreatment of poplar wood chips with Phanerochaete chrysosporium KCCM 34740 for 10days resulted in a some increase in screened yield (by 2%). According to increase of fungal incubation time, decrease in freeness (CSF) and increase in WRY were observed And bio-kraft pulping also led to improvement of physical properties of handsheet. As a result of bio-beating effect, we expect the saving of chemicals in kraft cooking process and energy consumption in beating process.

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Screening of Wood-Rotting Fungi for Efficient Decolorization of Draft Pulp Bleaching Effluents

  • Lee, Seon-Ho
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.5
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    • pp.95-100
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    • 1999
  • In order to find fungi having high treatment activity of kraft pulp bleaching (E1) effluent without any additional nutrietns, 124 strains of white-rot fungi were isolated from decayed wood samples. The author isolated five fungi(KS-62, MZ-400 , YK-719, YK-472 and Phanerochaete sordida YK-624) having high-decolorization activity of the E1 effluent. Particularly, the fugus KS-62 show the high effect of the decolorization and the degradation of the chlorinated lignin in the E1 , effluent compared with Coriolus versicolor and Phanerochaete chrysosporium.

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Biodegradation of Kraft Lignins by White-Rot Fungi(I) -Lignin from Pitch Pine- (백색부후균에 의한 크라프트 리그닌의 분해(I) -리기다소나무 리그닌-)

  • 김명길;안원영
    • Journal of Korea Foresty Energy
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    • v.17 no.1
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    • pp.56-70
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    • 1998
  • This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.

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