• Applied Biological Chemistry
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• 제47권3호
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• pp.287-292
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• 2004

Phellinus igniarius 26005에 의한 lignin peroxidase의 생산 조건의 최적화를 검토하였다. Lignin peroxidase는 진탕배양보다는 정치배양에서 생산성이 높았으며 최적 생산배지는 malt extract 1 g, yeast extract 0.4 g, glucose 0.4 g, 증류수 100 ml이었다. 효소 생산 유도 물질, Tween 80을 0.005% 수준으로 첨가하였을 때 가장 높은 효소 생산성을 보였으며 veratryl alcohol도 0.4 mM 수준에서 생산성 향상을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.179-188
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• 2017

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

• 한국균학회지
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• 제40권2호
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• pp.98-103
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• 2012

본 연구에서는 백색부후균 중 줄버섯, 단색털구름버섯, 산느타리 및 유관버섯 등의 균사체를 이용하여 congo red, amaranth, orange G 및 methylene blue 등의 합성염료 탈색에 관한 실험을 수행하였다. 실험 결과 줄버섯과 단색털구름버섯은 congo red가 함유된 고체와 액체배지에서 이들 염료를 93~95% 탈색하였으며 amaranth는 약 80%, orange G는 62~70% 탈색시키는 것으로 나타났으나 유관버섯에 의한 3종류의 염료 탈색율은 30% 내외로 매우 낮았다. congo red, amaranth 및 orange G 등 각각의 염료가 첨가된 배지에서의 염료 탈색율은 이들 배지에서 배양한 균사체의 생장과 상관관계가 있는 것으로 나타났다. 그러나 모든 공시 균주는 methylene blue가 함유된 고체와 액체배지에서 methylene blue를 효과적으로 탈색하지 못하는 것으로 나타났다. 공시된 백색부후균의 액체 배지에서의 리그닌 분해효소 생산을 탐색하기 위해 1%의 나프탈렌이 첨가된 PDB 배지에 공시균을 10일 간 배양 후 효소의 종류와 양을 분석한 결과 모든 공시균은 laccase, lignin peroxidase 그리고 manganese peroxidase 등의 효소를 생산하는 것으로 확인되었으며 공시균주 중 줄버섯이 리그닌 분해 효소의 생산이 가장 왕성한 것으로 나타났다.

• Forest Science and Technology
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• 제13권4호
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• pp.158-163
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• 2017

We examined the activities of lignin-degrading enzymes of the mycelium of cauliflower mushroom (Sparassis latifolia). Three different strains of S. latifolia collected from several sites in Korea and one crossbred strain were cultured on potato dextrose broth (PDB) and Kirk's medium in order to study the activities of their ligninolytic enzymes. Mycelial growth reached maximum levels between 14 and 21 days after inoculation and pH increased by 0.12 units over 35 days. Laccase activity began increasing after 14 days on both types of media. Manganese peroxidase (MnP) activity followed a trend similar to that of laccase on Kirk's medium, but not on PDB. The activity of lignin peroxidase (LiP) differed from that of other enzymes; its activity decreased by half after 14 days on PDB but remained constant on Kirk's medium over 35 days. The total protein concentration increased considerably after 14 days and peaked at 21 days on PDB. A similar maximum was attained on Kirk's medium. In contrast, the residual glucose increased rapidly at 14 days on Kirk's medium, while increasing gradually up to 28 days on PDB. This study indicates that S. latifolia is more similar to white rot fungi than to other brown rot fungi.

• 한국버섯학회지
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• 제17권4호
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• pp.247-254
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• 2019

흰구름버섯 (Trametes hirsuta)의 균사체는 CR, CV, RBBR 등 방향족 염료가 함유된 고체와 액체 배지에서 이들 염료를 효과적으로 탈색하였으나 MB의 탈색은 저조하였다. 각각 CR, MB, CV 및 RBBR 등 4종류의 염료가 함유된 액체배지에서 흰구름버섯의 균사체를 10일 간 배양했을 때 laccase, LiP, MnP 등 세 종류의 효소를 모두 생산하였으며 이들 효소 중 laccase의 활성도가 가장 높았으며 LiP와 MnP의 활성도laccase에 비해 낮았다. 따라서 흰구름버섯 균사체에 의한 방향족 염료의 탈색에는 laccase가 주로 사용되고 LiP나 MnP는 보조적인 역할을 하는 것으로 사료된다. 또한 비스페놀 A가 0, 25, 50, 100, 200 ppm의 농도로 함유된 PDA 배지에 균사체를 접종하여 배양한 결과 비스페놀 A의 농도가 증가함에 따라 균사체의 생장은 농도 의존적으로 저해되는 것으로 나타났다. 또한 비스페놀 A가 100 ppm 함유된 YMG 액체배지에 균사체를 접종하고 비스페놀 A의 분해율을 측정한 결과 배양 12시간 후 72.3%, 배양 24시간 후 95.3%, 그리고 배양 36시간 후에는 100% 분해된 것으로 나타났다. 따라서 본 연구 결과는 우리나라의 산업 활동 과정에서 생산되고 자연계로 배출되어 생물체에 큰 피해를 주는 합성염료와 내분비계 장애물질인 비스페놀 A를 친환경적으로 제거할 수 있는 기술의 개발에 도움이 될 수 있을 것으로 사료된다.

• Journal of Microbiology
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• 제43권6호
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• pp.555-560
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• 2005

Laccase is one of the ligninolytic enzymes of white rot fungus Trametes versicolor 951022, a strain first isolated in Korea. This laccase was purified 209-fold from culture fluid with a yield of $6.2\%$ using ethanol precipitation, DEAE-Sepharose, Phenyl-Sepharose, and Sephadex G-100 chromatography. T. versicolor 951022 excretes a single monomeric laccase showing a high specific activity of 91,443 U/mg for 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as a substrate. The enzyme has a molecular mass of approximately 97 kDa as determined by SDS-PAGE, which is larger than those of other laccases reported. It exhibits high enzyme activity over broad pH and temperature ranges with optimum activity at pH 3.0 and a temperature of $50^{\circ}C$. The $K_m$ value of the enzyme for substrate ABTS is $12.8{\mu}M$ and its corresponding $V_{max}$ value is 8125.4 U/mg. The specific activity and substrate affinity of this laccase are higher than those of other white rot fungi, therefore, it may be potentially useful for industrial purposes.

• Asian-Australasian Journal of Animal Sciences
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• 제23권5호
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• pp.680-692
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• 2010

This paper gives an overview of the availability, nutritive quality, and possible strategies to improve the utilization of rice straw as a feed ingredient for ruminants. Approximately 80% of the rice in the world is grown by small-scale farmers in developing countries, including South East Asia. The large amount of rice straw as a by-product of the rice production is mainly used as a source of feed for ruminant livestock. Rice straw is rich in polysaccharides and has a high lignin and silica content, limiting voluntary intake and reducing degradability by ruminal microorganisms. Several methods to improve the utilization of rice straw by ruminants have been investigated in the past. However, some physical treatments are not practical because of the requirement for machinery or treatments are not economical feasible for the farmers. Chemical treatments, such as NaOH, $NH_3$ or urea, currently seem to be more practical for onfarm use. Alternative treatments to improve the nutritive value of rice straw are the use of ligninolytic fungi (white-rot fungi), with their extracellular ligninolytic enzymes, or specific enzymes degrading cellulose and/or hemicellulose. The use of fungi or enzyme treatments is expected to be a more practical and environmental-friendly approach for enhancing the nutritive value of rice straw and can be costeffective in the future. Using fungi and enzymes might be combined with the more classical chemical or physical treatments. However, available data on using fungi and enzymes for improving the quality of rice straw are relatively scarce.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1421-1430
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• 2009

With an aim to evaluate dye decolorization by white rot fungus on natural living conditions, reproducing by solid-state fermentation, the process of triphenylmethane dyes decolorization using the white rot fungus P. ostreatus BP, cultivated on rice straw solid-state medium, has been demonstrated. Three typical dyes, including malachite green, bromophenol blue, and crystal violet, were almost completely decolorized by the fungus after 9 days of incubation. During the process of dye decolorization, the activities of enzyme secreted by the fungus, and the contents of soluble components, such as phenolic compounds, protein, and sugar, changed regularly. The fungus could produce ligninolytic, cellulolytic, and hemicellulolytic enzymes and laccase was the most dominant enzyme in solid-state medium. Laccase, laccase isoenzyme, and the laccase mediator could explain the decolorization of malachite green, bromophenol blue, and crystal violet by the fungus in solid-state medium, respectively. It is worth noting that the presence of the water-soluble phenolic compounds could stimulate the growth of fungus, enhance the production of laccase, and accelerate dye decolorization.

• Journal of the Korean Wood Science and Technology
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• 제21권3호
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• pp.87-93
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• 1993

This study was undertaken to investigate the characteristics and the productivities of lignin olytic enzymes: laccase (Lac) and Mn-dependent peroxidase (MnP) from Coriolus versicolor and Lentinus edodes respectively. Enzymes were isolated from cultural filterates and purified according to the standard methods. These enzymes showed one band in SDS-PAGE and their molecular weights were found 62,000 and 45,000 dalton respectively. Polyclonal antibodies against Lac and MnP were raised against mouse. In the ELISA (enzyme-linked immunosorbent assay), Lac and MnP-antiserum produced a strong positive reaction with Lac and MnP antigen($A_{405}$=2.50 and 3.53 respectively). The sera to negative (S/N) ratio was determined by the dividing the mean absorbance of antibodies by the corresponding diluted samples from normal mouse serum. The sera produced showed 2 times more positive reaction in S/N ratio than negative sera.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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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.