• Title/Summary/Keyword: lignin peroxidase

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Production of Mn-Dependent Peroxidase from Bjerkandera fumosa and Its Enzyme Characterization

  • Jarosz-Wilkolazka, Anna;Luterek, Jolanta;Malarczyk, Elzbieta;Leonowicz, Andrzej;Cho, Hee-Yeon;Shin, Soo-Jeong;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.2
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    • pp.85-95
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    • 2007
  • Manganese dependent peroxidase (MnP) is the most ubiquitous enzyme produced by white-rot fungi, MnP is known to be involved in lignin degradation, biobleaching and oxidation of hazardous organopollutants. Bjerkandera fumosa is a nitrogen-unregulated white-rot fungus, which produces high amounts of MnP in the excess of N-nutrients due to increased biomass yield. The objective of this study was to optimize the MnP production in N-sufficient cultures by varying different physiological factors such as Mn concentration, culture pH, and incubation temperature. The growth of fungus was optimal in pH 4.5 at $30^{\circ}C$, $N_2$-unregulated white-rot fungus produces high amounts of MnP in the excess N-nutrients. The fungus produced the highest level of MnP (up to $1000U/{\ell}$) with $0.25g/{\ell}$ asparagine and $1g/{\ell}$ $NH_4Cl$ as N source at 1.5 mM $MnCl_2$ concentration, pH value of 4.5 at $30^{\circ}C$. Purification of MnP revealed the existence of two isoforms: MnPl and MnP2. The molecular masses of the purified MnPl and MnP2 were in the same range of 42~45 kDa. These isoforms of B. fumosa strictly require Mn to oxidize phenolic substrates. Concerned to kinetic constants of B. fumosa MnPs, B. fumosa has similar Km value and Vmax compared to the other white-rot fungi.

Effect of $Zn^{2+}$ and Ferulic Acid on Laccase and Manganese Peroxidase Production by Funalia trogii (Funalia trogii에 의한 Laccase와 Manganese Peroxidase의 생산시 $Zn^{2+}$ 및 Ferulic Acid가 미치는 영향)

  • Park, Chul-Hwan;Han, Eun-Jung;Lee, Byung-Hwan;Lee, Jin-Won;Kim, Sang-Yong
    • KSBB Journal
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    • v.21 no.2
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    • pp.85-89
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    • 2006
  • Typical property of the white-rot fungi is their ability to degrade lignin and other aromatic compounds with non-specific extracellular enzyme. In this work, the modification of the strain(Funalia trogii ATCC 200800) and the culture condition was performed to enhance enzyme productivity. Single cell was separated by the protoplasts formation and several putative laccase and manganese peroxidase inducers were tested. By adopting the modified strain, enzyme productivity increased comparing with that of the original strain. Extracellular enzyme formation was highly stimulated by the addition of copper and various aromatic compounds in the glucose-based culture medium.

Production of manganese peroxidase from Pleurotus ostreatus using a rotary draft tube bioreactor (RTB) and characterization of its activity (회전식 통풍관 생물반응기 사용에 따른 느타리균의 manganese peroxidase 생산 및 특성)

  • Ha, Hyo-Cheol
    • 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.

Industrial applications and characteristics of lignocellulolytic enzymes in Basidiomycetous fungi (담자균류 목질섬유소 분해효소의 특성과 산업적 이용)

  • Lim, Sun-Hwa;Kang, Hee-Wan
    • 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.

Multifarious activities of cellulose degrading bacteria from Koala (Phascolarctos cinereus) faeces

  • Singh, Surender;Thavamani, Palanisami;Megharaj, Mallavarapu;Naidu, Ravi
    • Journal of Animal Science and Technology
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    • v.57 no.7
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    • pp.23.1-23.6
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    • 2015
  • Cellulose degrading bacteria from koala faeces were isolated using caboxymethylcellulose-Congo red agar, screened in vitro for different hydrolytic enzyme activities and phylogenetically characterized using molecular tools. Bacillus sp. and Pseudomonas sp. were the most prominent bacteria from koala faeces. The isolates demonstrated good xylanase, amylase, lipase, protease, tannase and lignin peroxidase activities apart from endoglucanase activity. Furthermore many isolates grew in the presence of phenanthrene, indicating their probable application for bioremediation. Potential isolates can be exploited further for industrial enzyme production or in bioremediation of contaminated sites.

Production of Lignocellulytic Enzymes from Spent Mushroom Compost of Pleurotus eryngii (큰느타리버섯 수확 후 배지로부터 리그닌섬유소분해효소 생산)

  • Lim, Sun-Hwa;Kim, Jong-Kun;Lee, Yun-Hae;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.152-158
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    • 2012
  • The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.

Optimal Extraction and Characteristics of Lignocellulytic Enzymes from Various Spent Mushroom Composts (다양한 버섯 수확 후 배지로부터 목질섬유소 분해효소의 최적 추출 및 특성)

  • Lim, Sun Hwa;Lee, Yun Hae;Kang, Hee Wan
    • The Korean Journal of Mycology
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    • v.41 no.3
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    • pp.160-166
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    • 2013
  • Recovery of ${\alpha}$-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) from spent mushroom composts (SMCs) of Pleurotus cornucopiae, Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceum, Lyophyllum ulmarium, Agrocybe cylindracea, Lentinus lepideus, and Flammulina velvtipes were investigated using different extraction buffers. The maximum recovery of the enzymes was mostly detected in SMC extracts with tap water and 0.25% Triton X-100 by shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. The xylanase (152 U/g) and laccase (8.1 U/g) activities were the highest in SMC extracts from F. velvtipes and P. eryngii. In addition, high enzymatic activities of ${\alpha}$-amylase (3.6 U/g) and cellulase (3.4 U/g) was detected in SMC extract of A. cylindracea. Futhermore, cellulase and laccase activities of SMCE from P. eryngii were compared to commercial enzymes.

The Effects of Phenolic Radical Carriers on the Enzymatic. Bleaching of Kraft Pulp (Kraft펄프의 효소표백반응에 미치는 페놀라디칼 전달체의 영향)

  • 류근갑
    • KSBB Journal
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    • v.10 no.2
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    • pp.183-190
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    • 1995
  • The effects of enzymatic pretreatments on the bleaching of kraft pulp were studied. The kappa number of pulp samples which represents the lignin content of pulp decreased by 25.2% by the pretreatments with xylanase(EC 3.2.1.8, Pulpzyme HB) while it decreased by 13.7% without enzyme pretreatments after the extraction of the pretreated pulp samples in 1N NaOH. To enhance the effects of enzymatic pretreatment on the bleaching of kraft pulp, phenols were used as radical carriers with the simultaneous use of peroxidase(EC 1.11.1.7, Novozyme 502), $H_2O_2$, and xylanase. Guaiacol (1mM) was most effective by decreasing the kappa number by 29.6% when a low initial concentration of $H_2O_2$ (0.1mM) was used. The use of either a higher initial concentration of $H_2O_2$ or phenols lacking electron donating substituents such as phenol and p-chloyophenol, however, decreased the efficiency of enzymatic pretreatment indicating that the production rate and the stability of phenolic radicals are important parameters.

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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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Biodegradation and Saccharification of Wood Chips of Pinus strobus and Liriodendron tulipifera by White Rot Fungi

  • Hwang, Soon-Seok;Lee, Sung-Jae;Kim, Hee-Kyu;Ka, Jong-Ok;Kim, Kyu-Joong;Song, Hong-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1819-1825
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    • 2008
  • Degradation and glucose production from wood chips of white pine (Pinus strobus) and tulip tree (Liriodendron tulipifera) by several white rot fungi were investigated. The highest weight losses from 4 g of wood chips of P. strobus and L. tulipifera by the fungal degradation on yeast extract-malt extract-glucose agar medium were 38% of Irpex lacteus and 93.7% of Trametes versicolor MrP 1 after 90 days, respectively. When 4 g of wood chips of P. strobus and L. tulipifera biodegraded for 30 days were treated with cellulase, glucose was recovered at the highest values of 106 mg/g degraded wood by I. lacteus and 450 mg/g degraded wood by T. versicolor. The weight loss of 10 g of wood chip of L. tulipifera by T. versicolor on the nutrient non-added agar under the nonsterile conditions was 35% during 7 weeks of incubation, and the cumulative amount of glucose produced during this period was 239 mg without cellulase treatment. The activities of ligninolytic enzymes (lignin peroxidase, manganese peroxidase, and laccase) of fungi tested did not show a high correlation with degradation of the wood chips and subsequent glucose formation. These results suggest that the selection of proper wood species and fungal strain and optimization of glucose recovery are all necessary for the fungal pretreatment of woody biomass as a carbon substrate.