• Title/Summary/Keyword: laccase3

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Identification and Characterization of Ligninolytic Enzyme by Serratia marcescens HY-5 isolated from the Gut of Insect

  • Kim, Gi-Deok;Sin, Dong-Ha;Son, Gwang-Hui;Park, Ho-Yong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.473-476
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    • 2002
  • A lignin degradation bacteria, symbiotic bacteria was isolated from the gut of Sympetrum depressiusculum and tested for its lignin degrading activity using lignin model compounds and related aromatic compounds. The strain was identified as Serratia marcescens HY-5 based on the 165 rDNA, cellular fatty acid composition, biochemical and physiological characteristics. S. marcescens showed 40-50% lignin degrading activity in the media that contained vaillin, guaiacol and dealkaline lignin. S. marcescens showed three ligninase activities [Jaccase, lignin peroxidase(LiP) and Manganase peroxidase(MnP)]. Addition of dealkaline lignin to the basal media increased about 6fold of laccase activity. Vanillic acid or vanillin increase 1.3fold of MnP activity and p-coumaric acid increased 12fold of LiP activity which added to the basal medium.

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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.

Simultaneous Degradation of Polycyclic Aromatic Hydrocarbons by Attractive Ligninolytic Enzymes from Phlebia brevispora KUC9045

  • Lee, Aslan Hwanhwi;Lee, Hanbyul;Kim, Jae-Jin
    • Korean Journal of Environmental Biology
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    • v.34 no.3
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    • pp.201-207
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    • 2016
  • The hazards associated with the polycyclic aromatic hydrocarbons (PAHs) are known to be recalcitrant by their structure, but white rot fungi are capable of degrading recalcitrant organic compounds. Phlebia brevispora KUC9045 isolated from Korea was investigated its efficiency of degradation of four PAHs, such as phenanthrene, anthracne, fluoranthene, and pyrene. And the species secreted extracellular laccase and MnP (Manganese dependent peroxidase) during degradation. P. brevispora KUC9045 demonstrated effective degradation rates of phenanthrene (66.3%), anthracene (67.4%), fluoranthene (61.6%), and pyrene (63.3%), respectively. For enhancement of degradation rates of PAHs by the species, Remazol Brilliant Blue R (RBBR) was preferentially supplemented to induce ligninolytic enzymes. The biodegradation rates of the three PAHs including phenanthrene, fluoranthene, and pyrene were improved as higher concentration of Remazol Brilliant Blue R was supplemented. However, anthracene was degraded with the highest rate among four PAHs after two weeks of the incubation without RBBR addition. According to the previous study, RBBR can be clearly decolorized by P. brevispora KUC9045. Hence, the present study demonstrates simultaneous degradation of dye and PAHs by the white rot fungus. And it is considered that the ligninolytic enzymes are closely related with the degradation. In addition, it indicated that dye waste water might be used to induce ligninolytic enzymes for effective degradation of PAHs.

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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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.

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.

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.

Trametes sp. CJ-105에 의한 염료의 색도제거

  • Kim, Hyun-Soo;Oh, Kwang-Keun;Lee, Cheol-Woo;Lee, Jae-Heung;Jeon, Yeong-Joong
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.630-635
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    • 1997
  • Decolorization of congo red, methyl orange, poly R478, remazol brilliant blue R and crystal violet by white-rot fungus Trametes sp. CJ-105, isolated in Korea, was investigated. Remazol blue and methyl orange were almost completely decolorized after 2 days of culture, but congo red, crystal violet and poly R478 were decolorized by about 80%, 40% and 30% after 10 days of culture, respectively. As a result of determination of cell mass and enzyme activity, it was shown that color removal efficiency was related to cell mass and enzyme activity, and also found that only laccase (E.C.1.10.3.2) activity was existed in the culture broth. The decolorization ratios of remazol blue in the concentrations of 100ppm to 3, 000 ppm were 85% and above after 2 days of culture. In this study, we found that white-rot fungus, Trametes sp. CJ-105, was effective in decolorizing a wide range of structurally different synthetic dyes.

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Decolorization of Dyehouse Effluent and Biodegradation of Congo Red by Bacillus thuringiensis RUN1

  • Olukanni, O.D.;Osuntoki, A.A.;Awotula, A.O.;Kalyani, D.C.;Gbenle, G.O.;Govindwar, S.P.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.843-849
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    • 2013
  • A dye-decolorizing bacterium was isolated from a soil sample and identified as Bacillus thuringiensis using 16S rRNA sequencing. The bacterium was able to decolorize three different textile dyes, namely, Reactive blue 13, Reactive red 58, and Reactive yellow 42, and a real dyehouse effluent up to 80-95% within 6 h. Some non-textile industrially important dyes were also decolorized to different extents. Fourier transform infrared spectroscopy and gas chromatography-mass spectrometer analysis of the ethyl acetate extract of Congo red dye and its metabolites showed that the bacterium could degrade it by the asymmetric cleavage of the azo bonds to yield sodium (4-amino-3-diazenylnaphthalene-1-sulfonate) and phenylbenzene. Sodium (4-amino-3-diazenylnaphthalene-1-sulfonate) was further oxidized by the ortho-cleavage pathway to yield 2-(1-amino-2-diazenyl-2-formylvinyl) benzoic acid. There was induction of the activities of laccase and azoreductase during the decolorization of Congo red, which suggests their probable role in the biodegradation. B. thuringiensis was found to be versatile and could be used for industrial effluent biodegradation.

Water Absorption Properties and Biodegradability of Lignin/PVA Nanofibrous Webs (리그닌/PVA 나노섬유 웹의 수분 특성 및 생분해성 평가)

  • Song, Youjung;Lee, Eunsil;Lee, Seungsin
    • Journal of the Korean Society of Clothing and Textiles
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    • v.41 no.3
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    • pp.517-526
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    • 2017
  • The biodegradation and water absorption properties of lignin/poly(vinyl alcohol) (PVA) nanofibrous webs are investigated. Lignin/PVA nanofibrous webs containing 0, 50, and 85wt% of lignin were prepared via an electrospinning process to observe the effect of the lignin concentration on the biodegradability and water absorption properties of lignin/PVA nanofibrous webs. The morphology of the materials was examined by field emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). To understand the wetting behavior and hydrophilic nature of the electrospun lignin/PVA nanofibrous webs, the water absorbency, contact angle, and water uptake were examined. The enzymatic degradation of lignin/PVA nanofibrous webs was investigated using laccase by measuring total organic carbon (TOC) concentration over a course of 50 days. Water drops were absorbed immediately into all of the specimens. The water uptake of lignin/PVA nanofibrous webs increased as the amount of PVA in the lignin/PVA hybrid webs increased. The enzymatic degradation experiment indicated that the inherent biodegradability of lignin was retained after its transformation into nanofibers. Our findings imply that blending these two types of polymers is promising because it can lead to the development of a new range of multifunctional materials such as antimicrobial absorbent nanotextiles based on sustainable biopolymers.