• 한국미생물·생명공학회지
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• 제27권6호
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• pp.477-483
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• 1999

For Trametes sp. CJ-105, a kind of white-rot fungi which was collected from the mountain of Korea and was proven to be effective in decolorizing a wide range of structurally different synthetic dyes, the optimum conditions for mycelial growth and laccase(E.C. 1.10.3.2) production were investigated. Among various carbon sources, glucose showed the highest potential for the mycelial growth and laccase production, the optimum concentration being 2% glucose. For the nitrogen source, asparagine was good for the mycelial growth, while ammonium tartrate for laccase production(optimum concentration: 0.04%). The addition of thiamine and biotin increased both th emycelial growth and laccase production. When 2,5-xylidine was added as an inducer after the first day of culture, the production of alccase was seven-times higher than that in the absence of the inducer. The optimum pH and temperature conditions for laccase production by Trametes sp. CJ-105 were pH 5.0 and $25^{\circ}C$, respectively. In the 5L fermentation, the production of laccase reached a maximum of 340U/ml at the time when the ammonium ion was being rapidly depleted.

• Journal of the Korean Wood Science and Technology
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• 제47권2호
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• pp.210-220
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• 2019

This work aimed to evaluate the influence of culture conditions on laccase production in the co-culture of wood-rotting fungus with Trichoderma sp. The effects of infection extent, infection time, and culture filtrate of Trichoderma sp. on the laccase production by wood-rotting fungus in co-culture were examined. T. rubrum LKY-7 and T. longibrachiatum were selected as fungi which are effective in co-culture for laccase production. A significant increase in laccase activity was observed when T. rubrum LKY-7 was co-cultured with T. longibrachiatum in glucose-peptone liquid medium, yielding an increase of more than 5 times in laccase activity, as compared with control. Laccase production by T. rubrum LKY-7 during co-culturing was significantly influenced by the infection extent and the infection time of T. longibrachiatum. Maximal laccase activity was obtained when T. rubrum LKY-7 culture was infected by T. longibrachiatum after 3 days of cultivation at an inoculum size ratio of 0.5 to 1. The addition of culture filtrate or autoclaved mycelium of T. longibrachiatum to T. rubrum LKY-7 culture did not significantly enhance laccase production by T. rubrum LKY-7 as compared with control (mono cultures of T. rubrum LKY-7).

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1757-1764
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• 2013

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and $CuSO_4$ were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, $(NH_4)_2SO_4$ 1 g/l, $CuSO_4$ 0.51 g/l, Tween-20 1 g/l, $MgSO_4$ 1 g/l, and $KH_2PO_4$ 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

• 펄프종이기술
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• 제35권5호
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• pp.80-86
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• 2003

The wood-degrading fungus Trichophyton sp. LKY-7 (T. LKY-7) was immobilized in ca-alginate beads for laccase production and PCP bioremediation. The immobilized T. LKY-7 enabled the repeated use of this fungus for laccase production and produced high amount of laccase throughout 5 cycles incubation. As a laccase inducer. oak wood meal (Quercus variabilis) seemed to be effective laccase inducer for T. LKY-7, and the optimum addition amount was 1％ (W/W) in glucose-peptone medium. Bioremediation of pentachlorophenol by the immobilized T. LKY-7 reached an efficency of up to 90％ without toxic inhibition. The immobilized T. LKY-7 might thus be applicable for semicontinuous laccase production and bioremediation to serve inoculum for reactor system.

• 한국펄프종이공학회:학술대회논문집
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• 한국펄프종이공학회 2003년도 춘계학술발표논문집
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• pp.180-187
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• 2003

The lignin-degrading fungus Trichophyton sp. LKY-7 was immobilized in ca-alginate beads for laccase production and PCP remediation. The immobilized Trichopphyton sp. LKY-7 enabled the repeated use of this fungus for laccase production and produced high amount of laccase throughout 5 cycles incubation. As a laccase inducer oak wood meal(Quercus variabilis) seemed to be effective laccase inducer for Trichophyton sp. LKY-7, and the optimum addition amount was 1%(W/W) in glucose-peptone medium. Biotransformation of pentachlorophenol by immobilized Trichophyton sp. LKY-7 reached an efficency of up to 90% without toxic inhibition. Immobilized Trichophyton sp. LKY-7 might thus be applicable for semicontinuous laccase production and bioremediation to serve inoculum for reactor system.

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.228-234
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• 2006

장수버섯(Fomitella fraxinea)의 균사체 배양액으로부터 laccase를 생산하기 위한 최적 배양조건을 조사하였다. 복합 배지 중에서 MCM이 laccase의 생산에 가장 우수하였으며, laccase를 생산하기 위한 최적 배지조건은 탄소원, 질소원, 인산원 및 무기질원으로 각각 2% dextrose, 0.4% $(NH_4)_{2}HPO_4$, 0.05% $Na_{2}HPO_{4}{\cdot}7H_{2}O$ 및 0.05% KCl의 첨가에 의하여 효소의 생산이 가장 높았다. 따라서 E. fraxinea로 부터 laccase를 생산하기 위한 최적 배지조건은 2% dextrose, 0.4% $(NH_4)_{2}HPO_4$, 0.05% $Na_{2}HPO_{4}{\cdot}7H_{2}O$ 및 0.05% KCl 이다. 이상의 배지를 사용하여 배양온도 $25^{\circ}C$, 초기 pH 8.0에서 10일 동안 배양하였을 때 효소의 생산이 가장 증가함을 알 수 있었다. Staib agar plate 상에서 장수 버섯(Fomitella fraxinea) 균사체의 laccase활성여부를 확인할 수 있었으며 배양액 중의 효소 활성의 최적 pH와 온도는 pH 5.0과 $50^{\circ}C$ 이었다.

• Mycobiology
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• 제44권4호
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• pp.260-268
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• 2016

Regulation of alkaline-resistant laccase from Perenniporia tephropora KU-Alk4 was proved to be controlled by several factors. One important factor was the initial pH, which drove the fungus to produce different kinds of ligninolytic enzymes. P. tephropora KU-Alk4 could grow at pH 4.5, 7.0, and 8.0. The fungus produced laccase and MnP at pH 7.0, but only laccase at pH 8.0. The specific activity of laccase in the pH 8.0 culture was higher than that in the pH 7.0 culture. At pH 8.0, glucose was the best carbon source for laccase production but growth was better with lactose. Low concentrations of glucose at 0.1% to 1.0% enhanced laccase production, while concentrations over 1% gave contradictory results. Veratryl alcohol induced the production of laccase. A trace concentration of copper ions was required for laccase production. Biomass increased with an increasing rate of aeration of shaking flasks from 100 to 140 rpm; however, shaking at over 120 rpm decreased laccase quantity. Highest amount of laccase produced by KU-Alk4, 360 U/mL, was at pH 8.0 with 1% glucose and 0.2 mM copper sulfate, unshaken for the first 3 days, followed by addition of 0.85 mM veratryl alcohol and shaking at 120 rpm. The crude enzyme was significantly stable in alkaline pH 8.0~10.0 for 24 hr. After treating the pulp mill effluent with the KU-Alk4 system for 3 days, pH decreased from 9.6 to 6.8, with reduction of color and chemical oxygen demand at 83.2% and 81%, respectively. Laccase was detectable during the biotreatment process.

• 생명과학회지
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• 제25권6호
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• pp.673-679
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• 2015

본 연구에서는 재비늘버섯(Pholiota highlandensis)의 리그닌 분해 활성을 확인하고, laccase 생산을 위한 최적 배양 조건을 조사하였다. 재비늘버섯 균사체로부터 laccase를 생산하기 위한 배지조건을 조사한 결과, 다양한 합성 배지 중에서 Coriolus versicolor 배지(2% dextrose, 0.4% peptone, 0.6% yeast extract, 0.046% KH₂PO₄, 0.1% K₂HPO₄, 0.05% MgSO₄·7H₂O)가 가장 높은 laccase 활성을 나타내었다. Laccase생산에 대한 배양 조건을 최적화하기 위하여 Coriolus versicolor 배지의 조성 중에서 탄소원, 질소원, 인산원, 무기염에 대한 영향을 조사하였다. 탄소원, 질소원, 인산원, 무기염은 각각 2% fructose, 0.4% peptone 및 0.6% yeast extract, 0.05% NaH₂PO₄, 0.05% MgSO₄·7H₂O의 경우에 가장 높은 효소활성을 나타내었다. 본 균주는 몇 가지 방향족 화합물에 의해 laccase 생산이 유도되었고, guaiacol을 첨가할 경우 25℃에서 11일 동안 배양하였을 때 효소의 활성이 최대치(114.1 U/ml)에 도달하였다. 또한 본 균주의 laccase 생산을 위한 최적 pH와 온도는 각각 8.0과 35℃를 나타내었다. 최적 조건에서 배양된 균사체 배양 상등액을 Native-PAGE로 전기영동한 후 ABTS를 기질로 사용하여 활성염색을 수행한 결과, 약 90 kDa 부근에서 laccase 활성을 가지는 1개의 밴드를 확인하였다.

• 한국미생물·생명공학회지
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• 제13권1호
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• pp.65-70
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• 1985

The production media, partial purification and the enzymatic characteristics of laccase from Pleurotus ostreatus were investigated. Among various media tried the double strength onion media showed much higher production of laccase compared to glucose and/or CMC added media. The laccase from Pleurotus ostreatus has the optimum pH of 5.94 for the activity and appears to be stable at relatively broad pH spectrum (4.7-8.7). The experiments on the temperature stability shows that above 90% activity could be preserved after holding at $30^{\circ}C$ for 40 min., 60% activity was remained after 40 min. at $50^{\circ}C$. The Km value of the laccase from Pleurotus ostreatus was estimated to be 3.209mM and the molecular weight of laccase was estimated to be 55,000 by SDS-Polyacrylamide gel electrophoresis.

• 한국미생물·생명공학회지
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• 제14권2호
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• pp.139-143
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• 1986

The production media and enzymatic characteristics of laccase from Ganoderma lucidum was investigated. Potato dextrose yeast extract media was proved to be the best for laccase production. The enzyme has optimum pH of 6.45km value of 6.71 mM and appeared to be stable at wide pH range. The enzyme was inactivated partially by methanol and ethanol and totally by sodium azide but not at all by acetone. Also the enzyme purification was performed and the data is given.