• Title/Summary/Keyword: laccase

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Mycelial growth and wood decaying enzymatic activity analysis by various addition rates of oak powder in the liquid spawn of Lentinula edodes (참나무분 첨가에 따른 표고 액체종균의 균체생산 및 효소 활성)

  • Kim, Jeong-Han;Kang, Young-Ju;Baek, Il-Sun;Jeoung, Yun-Kyeoung;Lee, Yong-Seon;Cho, Hae-Seok;Lee, Young-Soon
    • Journal of Mushroom
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    • v.16 no.2
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    • pp.74-78
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    • 2018
  • This study was carried out to establish a suitable method for liquid spawn production from Lentinula edodes. The optimum production of liquid spawn (OLS) was achieved using soybean meal medium (SMM) with 0.3% of 850 um oak powder and 10-day incubation period and 0.6 vvm aeration volume. OLS showed activities of laccase on ABTS agar plate and carboxymethyl cellulase (CM-cellulase) on CMC agar plate. In case of liquid spawn, fruiting-body development period was delayed approximately 1 day compared to that of sawdust spawn, however, the yield of 153 g per 1.2 kg polypropylene bag was similar to that of sawdust spawn.

Decolorization of Synthetic Dyes and Ligninolytic Enzymes Production by White Rot Fungi (백색부후균에 의한 합성염료의 탈색과 리그닌분해 효소의 생산)

  • Gu, Bon-Joon;Kim, Min-Sik;Kim, Yin-Man;Kim, Seon-Woong;Choi, Won-Hyeok;Lee, Mi-Hwa;Cho, Hae-Jin;Lee, Tae-Soo
    • 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.

Decolorization Efficiency of Different Dyes by Extract from Spent Mushroom Substrates of Pleurotus eryngii (큰느타리(Pleurotus eryngii)의 수확 후 배지추출물을 이용한 다양한 염료의 탈색효과)

  • Lim, Seon Hwa;Kwak, A Min;Min, Gyeong Jin;Kim, Sang Su;Lee, Sang Yeop;Kang, Hee Wan
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.213-218
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    • 2014
  • Water extract from spent mushroom substrates (SMS) of Pleurotus eryngii was utilized in decolorization of eight synthetic dyes and wastewater from a textile factory. High laccase activity was detected in the extract of P. eryngii (SMSE). The SMSE showed that decolorization rate was 34~93% after 24 h incubation without any mediator on eight dyes including Rit-blue and Rit-red used in fiber dyeing. Dye decolorization rate more than 90% was observed on bromophenol blue and remazol brilliant blue R (RBBR). Dye in textile wastewater was decolorized at room temperature after three days by addition of P. eryngii SMSE. The results suggest that biological decolorization of dyes using the P. eryngii SMSE can be used as environmental friendly materials.

Ergosterol Contents and Enzymatic Characteristics of Lentinula edodes During Culture and Fruiting Periods (표고 균주의 배양 기간과 자실체 발생 기간에 따른 에르고스테롤 변화와 효소적 특성)

  • Kim Myungkil;Yoon Kabhee;Bak Wonchull;Park Hyun;Choi Joonweon;Lee Jaewon;Lee Bonghun
    • Journal of Korea Foresty Energy
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    • v.23 no.2
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    • pp.21-28
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    • 2004
  • Three different strains of Lentinula edodes, Sanlim 5-Ho, Sanlim 6-Ho and Nongki 3-Ho, were cultured in the sawdust media of Mongolian oak(Quercu mongolica Fisch) for 90 days under dark and light conditions(each 30 days) and fruiting period(30 days). Weight loss of sawdust media was determined after fungal cultures and the contents of ergosterol in fungal mycelia were quantified by HPLC analysis followed by solvent extraction. Compared with the two other fungal strains$(8\%)$, weight loss of Sanlim 5-Ho was slightly lowered to $7\%$. The level of ergosterol content, a parameter for fungal growth, was continuously enhanced in Sanlim 5-Ho for dark and light incubation periods. However, Sanlim 6-Ho and Nongki 3-Ho recorded the maximized fungal growth under light condition. In fruiting periods the ergosterol contents were lowered in the three strains. Intra- and extracellular enzymes during cultural and fruiting periods were also characterized. The activity of Mn-peroxidase and laccase, which are characteristics enzymes for white rot fungi as lignin degrading enzymes, were determined as a high level overall the periods. As cellulose degrading indicators, the activity of CMCase, avicelase, xylanase and glucanase were detectable in initial incubation period.

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Fundamental Cultural Characteristics of Chlorociboria spp. Native to Korea (한국에 자생하는 녹청균류의 기초 배양 특성)

  • Jeon, Sung-Min;Ka, Kang-Hyeon;Wang, Eun-Jin;Ryoo, Rhim;Jang, Yeongseon
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.145-160
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    • 2018
  • Members of Chlorociboria are soft-rot ascomycetes that produce blue-green pigment. We investigated the growth characteristics of two Korean species of Chlorociboria, eight strains of Chlorociboria aeruginascens and Chlorociboria poutoensis, under various culture conditions (solid media, temperature, pH) and screened them for extracellular enzyme activity. Although the growth rate was slow, all tested strains of Chlorociboria spp. grew well on potato dextrose agar (PDA; 16.3~42.6 mm after 60 days) or Sabouraud dextrose agar (SDA), but not on malt extract agar (MEA). Compared with C. aeruginascens strains, C. poutoensis strains exhibited higher expression of blue-green pigments on both PDA and SDA media. The optimal temperature for mycelial growth was $20{\sim}25^{\circ}C$, and mycelial growth was lower at $30^{\circ}C$ than at $10^{\circ}C$. All strains tended to have increased mycelial growth as the incubation temperature increased in the range of 10 to $20^{\circ}C$. The optimal pH of potato dextrose broth (PDB) for mycelial growth varied according to the strain under static culture conditions. Maximum biomass production was obtained at pH 6.0 for NIFoS 579 ($114.3{\pm}5.1mg/60days$), but it maintained a stable pigment expression under a broad pH spectrum. The activities of both cellulase and laccase were observed in all tested strains of Chlorociboria spp. Enzyme activities of NIFoS 579 were remarkably higher than those of the other strains. From these results, we suggest that C. poutoensis NIFoS 579 is a potential candidate for use as a source of natural blue-green dye.

Biodegradation of 4-t-Octylphenol by Basidioradulum molare and Schizopora paradoxa and Estrogenecity Reduction of its Metabolites (옥틸페놀(4-t-Octylphenol)의 Basidioradulum molare와 Schizopora paradoxa에 의한 분해 및 에스트로겐성 저감효과)

  • Lee, Soo-Min;Ku, Bon-Wook;Lee, Jae-Won;Choi, Don-Ha;Jeung, Eui-Bae;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.4
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    • pp.27-35
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    • 2004
  • Recalcitrant 4-t-Octylphenol used as a surfactant was subjected to the biodegradation with wood rot fungi, Basidioradulum molare and Schizopora paradoxa. Two fungi were grown in the culture medium containing various concentrations of 4-t-Octylphenol in order to investigate their resistance against 4-t-octylphenol Schizopora paradoxa was reached to the full growth within 14 incubation days in the concentration of more than 200 ppm of 4-t-Octylphenol, while Basidioradulum molare showed the inhibitory mycelium growth as the concentration was increased Schizopora paradoxa and Basidioradulum molare biodegraded 95% and 36% of initial concentration of 4-t-Octylphenol at first incubation day, respectively. However, the biodegradation capability reached to more than 95% after 3 incubation days. During the biodegradation of 4-t-Octylphenol, the activity of manganese dependent peroxidase was induced by the addition of 4-t-Octylphenol in the culture medium of Schizopora paradoxa, but that of laccase was maximal before the addition. The reduction of estrogenecity was assayed by MCF-7 cell proliferation test and measurement of pS2 mRNA expression. The level of pS2 mRNA was decreased down to the level of baseline at first incubation day. Also, estrogenecity of 4-t-Ocrylphenol completely disappeared after treatment with supernatant by Schizopora paradoxa and Basidioradulum molare from first incubation day of culture down to the levels of vehicle.

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.

Biodegradation of Pyrene by the White Rot Fungus, Irpex lacteus

  • Hwang, Soon-Suk;Song, Hong-Gyu
    • 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.

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Identification of Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides via Proteomics and Quantitative Real-Time PCR

  • Choi, Yoon-E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.648-657
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    • 2008
  • In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

Degradation and Detoxification of Disperse Dye Scarlet RR by Galactomyces geotrichum MTCC 1360

  • Jadhav, S.U.;Ghodake, G.S.;Telke, A.A.;Tamboli, D.P.;Govindwar, S.P.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.409-415
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    • 2009
  • Galactomyces geotrichum MTCC 1360 degraded the Scarlet RR(100 mg/l) dye within 18 h, under shaking conditions(150 rpm) in malt yeast medium. The optimum pH and the temperature for decolorization were pH 12 and $50^{\circ}C$, respectively. Enzymatic studies revealed an induction of the enzymes, including flavin reductase during the initial stage and lignin peroxidase after complete decolorization of the dye. Decolorization of the dye was induced by the addition of $CaCO_3$ to the medium. EDTA had an inhibitory effect on the dye decolorization along with the laccase activity. The metabolites formed after complete decolorization were analyzed by UV-VIS, HPLC, and FTIR. The GC/MS identification of 3 H quinazolin-4-one, 2-ethylamino-acetamide, 1-chloro-4-nitro-benzene, N-(4-chloro-phenyl)-hydroxylamine, and 4-chloro-pheny-lamine as the final metabolites corroborated with the degradation of Scarlet RR. The phytotoxicity study revealed the nontoxic nature of the final metabolites. A possible degradation pathway is suggested to understand the mechanism used by G. geotrichum and thereby aiding development of technologies for the application of this organism to the cleaning-up of aquatic and terrestrial environments.