• Title/Summary/Keyword: Laccase

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Kinetic Evidence for the Interactive Inhibition of Laccase from Trametes versicolor by pH and Chloride

  • Raseda, Nasrin;Hong, Soonho;Kwon, O Yul;Ryu, Keungarp
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1673-1678
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    • 2014
  • The interactive inhibitory effects of pH and chloride on the catalysis of laccase from Trametes versicolor were investigated by studying the alteration of inhibition characteristics of sodium chloride at different pHs for the oxidation of 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). At pH 3.0, the addition of sodium chloride (50 mM) brought about a 40-fold increase in $K{_m}^{app}$ and a 4-fold decrease in $V_{max}{^{app}}$. As the pH increased to 7.0, the inhibitory effects of sodium chloride became significantly weakened. The mixed-inhibition mechanism was successfully used to quantitatively estimate the competitive and uncompetitive inhibition strengths by chloride at two different pHs (pH 3.0 and 6.0). At pH 3.0, the competitive inhibition constant, $K_i$, was 0.35 mM, whereas the uncompetitive inhibition constant, $K{_i}^{\prime}$, was 18.1 mM, indicating that the major cause of the laccase inhibition by chloride is due to the competitive inhibition step. At a higher pH of 6.0, where the inhibition of the laccase by hydroxide ions takes effect, the inhibition of the laccase by chloride diminished to a great extent, showing increased values of both the competitive inhibition constant ($K_i=23.7mM$) and uncompetitive inhibition constant ($K{_i}^{\prime}=324mM$). These kinetic results evidenced that the hydroxide anion and chloride share a common mechanism to inhibit the laccase activity.

Optimization of Laccase Production from Bacillus sp. PK4 through Statistical Design of Experiments

  • Rajeswari, Murugesan;Bhuvaneswari, Vembu
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.330-342
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    • 2017
  • Statistical design of experiments was employed to optimize the media composition for the production of laccase from Bacillus sp. PK4. In order to find the key ingredients for the best yield of enzyme production from the selected eleven variables viz yeast extract, glucose, zinc sulphate, copper sulphate, potassium chloride, magnesium sulphate, calcium chloride, ferrous sulphate, sodium chloride, potassium dihydrogen phosphate ($KH_2PO_4$) and dipotassium hydrogen phosphate ($K_2HPO_4$), Plackett-Burman design was applied. The $MgSO_4$, $FeSO_4$, and $CuSO_4$ showed positive estimate, and their concentration optimized further. The steepest ascent method and Box-Behnken method revealed that 1.5 mM $MgSO_4$, 0.33 g/l $FeSO_4$ and 1.41 mM $CuSO_4$ were optimal for the laccase production by Bacillus sp. PK4. This optimization strategy leads to enhancement of laccase production from 2.13 U/ml to 40.79 U/ml. Agro-wastes residues replace the carbon source glucose in the optimized media namely sugarcane bagasse, wheat bran, rice husk, and groundnut shell, among these groundnut shells (117 U/ml) was found to enhance the laccase production significantly. The laccase produced by Bacillus sp. PK4 was found to have the potential to degrade persistent organic pollutant benzo[a]pyrene.

Degradation of Endocrine Disrupting Chemicals by Laccase Transformant of Phlebia tremellosa (아교버섯 형질전환체를 이용한 내분비장애 물질의 분해)

  • Yeo, Su-Min;Kim, Myung-Kil;Choi, Hyoung T.
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.10-13
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    • 2008
  • Endocrine disrupting chemicals (EDCs) are hard to be degraded in nature, and are also accumulated in diverse organisms. They finally give negative effects to human through the food web. White rot fungi which have lignin-degrading enzymes have high potentials for degradation of recalcitrant compounds, and a white rot fungus, Phlebia tremellosa, isolated in Korea show good degrading activity against the endocrine disrupting phthalates. We have isolated a laccase cDNA which was involved in the degradation of EDCs, and constructed a laccase expression vector to use in the genetic transformation of P. tremellosa. The expression vector was stably integrated into the chromosomal DNAs and showed increased laccase activity in transformants. One of transformants showed not only increased degradation of several EDCs but also faster estrogenic decreasing activities generated by the EDCs.

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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Decolorization of Aromatic Dyes by White Rot Fungus Coriolus hirsutus (흰구름버섯(Coriolus hirsutus)에 의한 방향족 염료의 탈색)

  • Song, Yeon-Hong;Choi, Chul-Min;Kim, Chang-Jin;Shin, Kwang-Soo
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.252-256
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    • 1997
  • The white rot fungus Coriolus hiysutus decolorized several recalcitrant dyes. Four different types of dyes, including azo, triphenyl methane, heterocyclic, and polymeric dye, were treated by the mycelial preparation. Triphenyl methane dye, bromophenol blue lost over 95% of its color. Congo red and Poly R-478 were decolorized less than bromophenol blue, 57 and 55%, respectively. However, heterocyclic dye, methylene blue was not decolorized significantly and only red shift was observed. Extracellular laccase and peroxidase activities were appeared maximally in high level of dye decolorization media. In electrophoretic experiments, common active bands of laccase and peroxidase were found in all dye decolorized medium. These results indicated that the culture conditions which yield high levels of laccase and peroxidase activity lead to high levels of dye decolorization, and these two enzymes might be play an important roles in dye decolorization.

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Heterologous Expression and Characterization of a Laccase from Laccaria bicolor in Pichia pastoris and Arabidopsis thaliana

  • Wang, Bo;Yan, Ying;Xu, Jing;Fu, Xiaoyan;Han, Hongjuan;Gao, Jianjie;Li, Zhenjun;Wang, Lijuan;Tian, Yongsheng;Peng, Rihe;Yao, Quanhong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2057-2063
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    • 2018
  • Laccases can oxidize a variety of phenolic and non-phenolic substrates including synthetic dyes. In this research, a laccase gene Lcc9 from Laccaria bicolor was chemically synthesized and optimized to heterogeneous expression in Pichia pastoris and Arabidopsis thaliana. The properties of recombinant laccase expressed by P. pastoris were investigated. The laccase activity was optimal at 3.6 pH and $40^{\circ}C$. It exhibited $K_m$ and $V_{max}$ values of $0.565mmol\;l^{-1}$ and $1.51{\mu}mol\;l^{-1}\;min^{-1}$ for ABTS respectively. As compared with untransformed control plants, the laccase activity in crude extracts of transgenic lines exhibited a 5.4 to 12.4-fold increase. Both laccases expressed in transgenic P. pastoris or A. thaliana could decolorize crystal violet. These results indicated that L. bicolor laccase gene may be transgenically exploited in fungi or plants for dye decolorization.

Comparison of Lignocellulose degradation properties of Lentinula edodes varieties (표고(Lentinula edodes) 품종별 목질계 섬유소 분해효소 특성 비교)

  • Jeong, Sang-Wook;Jang, Eun-Kyoung;Choi, Seul-Ki;Seo, Kyoung-Sun;Jeong, Hee-Gyeong;Lee, Won-Ho;Ban, Seung-Eon
    • Journal of Mushroom
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    • v.20 no.1
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    • pp.29-33
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    • 2022
  • In this study, five different Lentinula edodes cultivar (Chamaram, Sanbaekhyang, Sanjo 713ho, Sanjo 715ho, Sanjo 718ho) were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in MEB medium, respectively. Chamaram and Sanjo 713ho decolorized RBBR rapidly in MEB medium within 3 and 5 days. The activities of manganese peroxidase (MnP) and laccase were determined on the MEB medium with and without lignin. Sanjo 713ho resulted the highest ligninolytic enzyme activities on incubation day 1, indicating of 1,213 U/mg of MnP activity and 1,421 U/mg of laccase activity.

Employing Laccase-Producing Aspergillus sydowii NYKA 510 as a Cathodic Biocatalyst in Self-Sufficient Lighting Microbial Fuel Cell

  • Abdallah, Yomna K.;Estevez, Alberto T.;Tantawy, Diaa El Deen M.;Ibraheem, Ahmad M.;Khalil, Neveen M.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1861-1872
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    • 2019
  • In the present work, we isolated and identified Aspergillus sydowii NYKA 510 as the most potent laccase producer. Its medium constituents were optimized to produce the highest possible amount of laccase, which was after 7 days at 31℃ and pH 5.2. Banana peel and peptone excelled in inducing laccase production at concentrations of 15.1 and 2.60 g/l, respectively. Addition of copper sulfate elevated enzyme yield to 145%. The fungus was employed in a microbial fuel cell (MFC). The best performance was obtained at 2000 Ω achieving 0.76 V, 380 mAm-2, 160 mWm-2, and 0.4 W. A project to design a self-sufficient lighting unit was implemented by employing a system of 2 sets of 4 MFCs each, connected in series, for electricity generation. A scanning electron microscopy image of A. sydowii NYKA 510 was utilized in algorithmic form generation equations for the design. The mixed patterning and patterned customized mass approach were developed by the authors and chosen for application in the design.

Overproduction of Laccase by the White-Rot Fungus Pleurotus ostreatus Using Apple Pomace as Inducer

  • Park, Young-Jin;Yoon, Dae-Eun;Kim, Hong-Il;Kwon, O-Chul;Yoo, Young-Bok;Kong, Won-Sik;Lee, Chang-Soo
    • Mycobiology
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    • v.42 no.2
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    • pp.193-197
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    • 2014
  • Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.