• Title/Summary/Keyword: Laccase Phanerochaete chrysosporium

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Improved Production of Ligninase and Laccase by Phanerochaete chrysosporium and Ceriporiopsis subvermispora (Phanerochaete chrysosporium과 Ceriporiopsis subvermispora 균주(菌株)의 Ligninase 및 Laccase 생산최적조건에 관한 연구(硏究))

  • Kang, An-Seok;Cha, Dong-Yeul;Kim, Kyung-Soo;Hong, In-Pyo;Croan, Suki C.;Yu, Seung-Hun
    • The Korean Journal of Mycology
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    • v.22 no.3
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    • pp.254-259
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    • 1994
  • The ever increasing demand for energy and the shortage of resources all over the world have generated interest in recycling renewable sources such as lignocelluloses which otherwise would go to waste and cause environmental pollution. Lignin is the incrustation material for cellulose and hemicellulose, therefore, cellulose and hemicellulose are not easily degraded. Recycling lignocellulosic wastes as agricultural material are benefit to everybody and everything. In order to improve ligninase and laccase production of Phanerochaete chrysosporium, BKM-F-1767 and Ceriporiopsis subvermispora, FP 90031-SP, were compared. The ligninase activity of P. chrysosporium was maximum on day 4.5 of shaking culture at 150 rpm 2.5 cm in a back and forth cycle. The laccase activity of C. subvermispora was maximum on day 5.5 for 2% malt extract+0.1% yeast extract+0.1% Tween 20+6 mM Benzyl alcohol culture medium at stationary state.

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Solid State Fermentation of Phanerochaete chrysosporium for Degradation and Saccharification of Lignocellulose (Phanerochaete chrysosporium의 고상발효를 통한 리그노셀룰로오즈 분해 및 당화)

  • Utomo, Romualdus N.C.;Lee, Eun-Kwang;Yun, Hyun Shik
    • KSBB Journal
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    • v.32 no.2
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    • pp.96-102
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    • 2017
  • The lignocellulose that is a major component of spent coffee ground was degraded and saccharified. To implement the spent coffee, after several pre-treatments, inoculation of Phanerochaete chrysosporium and solid-state fermentation were conducted. The optimal temperature of the enzymes (lignin peroxidase, manganese peroxidase, xylanase, laccase, and cellulase) for degradation of lignocellulose by P. chrysosporium was found. We also measured the maximum activity of enzymes (lignin peroxidase 0.15 IU/mL, manganese peroxidase 0.90 IU/mL, laccase 0.11 IU/mL, cellulase 5.87 IU/mL, carboxymethyl cellulase 9.52 IU/mL, xylanase 1.16 IU/mL) used for the process. As a result, 4.73 mg/mL of reduced sugar was obtained and 61.02% of lignin was degraded by solid state fermentation of P. chrysosporium on spent coffee ground.

Biodegradation of 4,5,6-Trichloroguaiacol by White Rot Fungi, Phanerochaete chrysosporium, Trametes versicolor, and Inonotus cuticularis (수질분해균(水質分解菌)에 의한 4,5,6-Trichloroguaiacol의 미생물분해(微生物分解))

  • Ahn, Sye-Hee;Choi, In-Gyu
    • 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.

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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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Autohydrolysis and Enzymatic Saccharification of Lignocellulosic Materials (IV) - Simultaneous Utilization of Laccase and Cellulase - (목질 재료의 자기가수분해 및 효소당화에 관한 연구 (IV) - Laccase 및 Cellulase의 동시 이용 가능성 -)

  • Cho, Nam-Seok;Lim, Chang-Suk;Lee, Jae-Sung
    • Journal of the Korean Wood Science and Technology
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    • v.17 no.3
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    • pp.52-60
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    • 1989
  • This study was carried out to know the possibility of simultaneous utilization of laccase from white-rot fungus with cellulase on enzymatic hydrolysis of cellulosic substrate from autohydrolyzed oak wood. Laccases from 3 white-rot fungi, Pleurotus ostreatus. Ganoderma lucidum, and Phanerochaete chrysosporium, were isolated, purified and measured their activities. The highest activity was shown in Pleurotus ostreatus and the lowest in Phanerochaete chrysosporium. Laccase from Pleurotus ostreatus has optimum pH of 5.94, Km value of 3.209 mM and appeared to be stable at relatively wide pH range, 4.7-8.72. Temperature stability showed that 60% activity was preserved after 40 minutes at $50^{\circ}C$. Laccase from Ganoderma lucidum reached to the maximum activity during 15-20 day incubation. This enzyme has optimum pH of 6.45, Km value of 6.71 mM and pH range of 5.0-9.0 for stabilization. 95% activity was preserved at $30^{\circ}C$ and 58% activity at $50^{\circ}C$. Concerned to the enzymatic hydrolysis of cellulosic substrate with both enzymes, cellulase and laccase, simultaneously, mixed culture filtrates and mycellium extracts were shown higher hydrolysis rates than those of Trichoderma viride. There were no significant differences in the extent of hydrolysis among various mixed culture filtrates and mycellium extracts.

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Estrogenic Reduction of Styrene Monomer Degraded by Phanerochaete chrysosporium KFRI 20742

  • Lee Jae-Won;Lee Soo-Min;Hong Eui-Ju;Jeung Eui-Bae;Kang Ha-Young;Kim Myung-Kil;Choi In-Gyu
    • 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.

Screening of Outstanding White Rot Fungi for Biodegradation of Organosolv Lignin by Decolorization of Remazol Brilliant Blue R and Ligninolytic Enzymes Systems (Remazol Brilliant Blue R 탈색능과 리그닌 분해 효소시스템을 이용한 유기용매 리그닌 생분해 우수 균주 선별)

  • Hong, Chang-Young;Kim, Ho-Yong;Jang, Soo-Kyeong;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.1
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    • pp.19-32
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    • 2013
  • In this study, outstanding white rot fungi for biodegradation of organosolv lignin were selected on the basis of their ligninolytic enzyme system. Fifteen white rot fungi were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in SSC and MEB medium, respectively. Six white rot fungi (Ceriporiopsis subvermispora, Ceriporia lacerate, Fomitopsis insularis, Phanerochaete chrysosporium, Polyporus brumalis, and Stereum hirsutum) decolorized RBBR rapidly in SSC medium within 3 days. The protein contents as well as the activities of manganese peroxidase (MnP) and laccase for 6 selected fungi were determined on the SSC medium with and without organosolv lignin. Interestingly, extracellular protein concentrations were determined to relative higher for S. hirsutum and P. chrysosporium in the presence of organosolv lignin than others. On the other hands, each fungus showed a different ligninolytic enzyme pattern. Among them, F. insularis resulted the highest ligninolytic enzyme activities on incubation day 6, indicating of 1,545 U/mg of MnP activity and 1,259 U/mg of laccase activity. In conclusion, $STH^*$ and FOI were considered as outstanding fungi for biodegradation of organosolv lignin, because $STH^*$ showed high extracellular protein contents and ligninolytic enzyme activities over all, and ligninolytic enzyme activities of FOI were the highest among white rot fungi used in this study.

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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Screening of White Rot Fungi with Selective Delignification Capacity for Biopulping (백색목재부후균중 Biopulping에 이용가능한 선택적 리그닌분해균의 스크리닝)

  • Lee, Jong-Kyu;Oh, Eun-Sung
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.144-152
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    • 1998
  • To obtain white rot fungi which have selective delignification capacity and can be used in biopulping processes, 94 different wood rotting fungi were screened and the capabilities of selected species were evaluated on deciduous and coniferous wood blocks. White rot fungi, first of all, were selected by simple enzyme tests, i.e., cellulase activity test; phenol oxidase activity test; laccase and peroxidase activity test. Most organisms that gave a positive Bavendamm gave a strongly positive laccase test with syringaldazine whereas most of those that gave a negative Bavendamm test also negative test for laccase and peroxidase, even if some exceptions were noted. Wood decay experiement were carried out to select fungal species with selective lignin-degrading ability by inoculating selected fungi to both wood blocks of Populus tomentiglandulosa and Larix leptolepis. After 12 weeks of incubation, weight losses, lignin losses, and morphological characteristics of the decayed wood were investigated. Almost all fungi tested caused 2 or more times of weight losses in P. tomentiglandulosa than in L. leptolepis, while no weight losses were detected from the un-inoculated wood blocks. Ceriporiopsis subvermispora and Phanerochaete chrysosporium were the best delignifiers for both hardwood and softwood. P. chrysosporium, however, was less effective than C. subvermispora. Bjerkandera adusta and two unidentified spp. caused delignification for only P. tomentiglandulosa. B. adusta caused simultaneous rot of all cell wall components, resulted in thinning of the secondary cell wall layers. Other fungi caused selective delignification resulting in the removal of lignin from middle lamella and separation of cells from each other.

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Biodegradation of Polycyclic Aromatic Hydrocarbons by White Rot Fungi (백색부후균을 이용한 다환방향족 탄화수소(PAHs) 의 분해)

  • 류원률;서윤수;장용근;조무환
    • KSBB Journal
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    • v.15 no.3
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    • pp.262-267
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    • 2000
  • The white rot fungi Phanerochaete chrysosporium(IFO 31249) Trametes sp and Pleurotus sp. were studied for their ability to degrade Polycyclic Aromatic Hydrocarbons(PAHs) using anthracene and pyrene as model compounds. The disapperarance anthracene and pyrene of from cultures of wild type strains. P chrysosporium Trametes sp. and Pleurotus sp was observed However the activities of ligninolytic enzymes were not detected in P chrysosporium cultures during degradation while ligninolytic enzymes were detected in both culture of Trametes sp. and Pleurotus sp. Therefore our results showed that PAHs was degraded under ligninolytic as well as nonligninolytic conditions. The results also indicate that lignin peroxidase(LiP) mananese peroxidase(MnP) and laccase are not essential for the biodegradation of PAHs by white rot fungi.

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