• KSBB Journal
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• 제17권1호
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• pp.14-19
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• 2002

본 연구의 목적은 회전생물반응기에 고정화한 P. chrysosporium IFO 31249를 이용하여 LiP 생산의 최적 조건을 조사하는 것이다. 회전생물반응기에서 batch culture시 최대 LiP 활성도는 300 U/L이었다. LiP 생산의 최적 조건은 드림의 회전 속도는 1 rpm, 담체 충진율은 20%이었다. MnP 생산을 위한 $MnSO_4$$\cdot$$H_2O$ 의 최적 농도는 50 ppm이었다. 그리고 산소의 충분한 공급은 LiP 생산의 가장 중요한 인자이었다. 또한 LiP 및 MnP의 생산에 있어 repeated-batch culture가 3번이나 가능하였다.

• Journal of the Korean Wood Science and Technology
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• 제35권2호
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• pp.85-95
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• 2007

Manganese dependent peroxidase (MnP) is the most ubiquitous enzyme produced by white-rot fungi, MnP is known to be involved in lignin degradation, biobleaching and oxidation of hazardous organopollutants. Bjerkandera fumosa is a nitrogen-unregulated white-rot fungus, which produces high amounts of MnP in the excess of N-nutrients due to increased biomass yield. The objective of this study was to optimize the MnP production in N-sufficient cultures by varying different physiological factors such as Mn concentration, culture pH, and incubation temperature. The growth of fungus was optimal in pH 4.5 at $30^{\circ}C$, $N_2$-unregulated white-rot fungus produces high amounts of MnP in the excess N-nutrients. The fungus produced the highest level of MnP (up to $1000U/{\ell}$) with $0.25g/{\ell}$ asparagine and $1g/{\ell}$ $NH_4Cl$ as N source at 1.5 mM $MnCl_2$ concentration, pH value of 4.5 at $30^{\circ}C$. Purification of MnP revealed the existence of two isoforms: MnPl and MnP2. The molecular masses of the purified MnPl and MnP2 were in the same range of 42~45 kDa. These isoforms of B. fumosa strictly require Mn to oxidize phenolic substrates. Concerned to kinetic constants of B. fumosa MnPs, B. fumosa has similar Km value and Vmax compared to the other white-rot fungi.

• Journal of Microbiology
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• 제45권3호
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• pp.213-218
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• 2007

Trametes versicolor has a lignin degrading enzyme system, which is also involved in the degradation of diverse recalcitrant compounds. Manganese-dependent peroxidase (MnP) is one of the lignin degrading enzymes in T. versicolor. In this study, a cDNA clone of a putative MnP-coding gene was cloned and transferred into an expression vector (pBARGPE1) carrying a phosphinothricin resistance gene (bar) as a selectable marker to yield the expression vector, pBARTvMnP2. Transformants were generated through genetic transformation using pBARTvMnP2. The genomic integration of the MnP clone was confirmed by PCR with bar-specific primers. One transformant showed higher enzyme activity than the recipient strain did, and was genetically stable even after 10 consecutive transfers on non-selective medium.

• 한국버섯학회지
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• 제10권2호
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• pp.74-82
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• 2012

백색부후균인 느타리버섯 60균주로부터 목재를 분해하는 리그닌-셀룰로오스 분해효소 생산 능력이 우수한 균주를 선발하였다. 그 결과 1, 2차 스크리닝을 통해 실험한 모든 균주에서 리그닌-셀룰로오스 분해효소를 생산하는 것을 확인하였다. 그러나, 아비셀 함유 평판배지에서 선발된 6개 균주의 경우 2차 스크리닝 아비셀-효모추출물-펩톤 액체배지에서 셀룰로오스 분해효소 활성이 낮은 것을 확인하였다. 자일란 분해효소의 경우 자일란-효모추출물-펩톤 액체배지에서 No. 6, No. 38균주에서 xylanse 1.0 U/ml 이상, 1,4-${\beta}$-xylosidase 0.15 U/ml 이상 생산되었다. RBBR 함유 평판배지에서 선발된 13개 균주를 가지고 글루코오즈-효모-펩톤 배지 조건하에서 리그닌 분해 효소의 생산 실험을 한 결과 락케이즈가 먼저 최대 활성(0.8~2.0U/ml)을 나타낸 뒤 Mn-dependent peroxidase가 최대활성(0.5~1.5U/ml)를 나타내었다. 한편, 글루코오즈-효모-펩톤 배지 조건하에서 실험한 모든 균주에서 lignin peroxidase는 생산되지 않았다. 특히 본 연구자는 이러한 스크리닝 방법에 의해 Mn-dependent peroxidase와 laccase 생산 능력이 높은 No. 42균주를 선발하였다.

• 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.

• Journal of Microbiology
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• 제42권1호
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• pp.37-41
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• 2004

The textile industry wastewater has been decolorized efficiently by the white rot fungus, Irpex lacteus, without adding any chemicals. The degree of the decolorization of the dye effluent by shaking or stationary cultures is 59 and 93%, respectively, on the 8th day. The higher level of manganese-dependent peroxidase (MnP) and non-specific peroxidase (NsP) was detected in stationary cultures than in the cultures shaken. Laccase activities were equivalent in both cultures and its level was not affected significantly by the culture duration. Neither lignin peroxidase (LiP) nor Remazol Brilliant Blue R oxidase (RBBR ox) was detected in both cultures. The absorbance of the dye effluent was significantly decreased by the stationary culture filtrate of 7 days in the absence of Mn (II) and veratryl alcohol. In the stationary culture filtrate, three or more additional peroxidase bands were detected by the zymogram analysis.

• 한국균학회지
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• 제26권1호통권84호
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• pp.8-15
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• 1998

우리 나라 자연 환경에서 분리 동정된 송곳니구름버섯(Irpex zonatus) BN2 균주의 리그닌분해효소활성과 아조(azo)계, 트리페닐메탄(triphenylmethane)계 및 헤테로싸이클릭(heterocyclic)계에 속하는 몇몇 염료의 탈색능을 조사하였다. 송곳니구름버섯 BN2 균주는 lignin peroxidase(LiP)와 veratryl alcohol oxidase(VAO)를 생산하지 않고 laccase와 manganese dependent peroxidase(MnP)를 생산했다. MnP는 배양 3일부터 생산되었으나 효소활성은 매우 낮았다. 반면 laccase는 배양 초기부터 지속적으로 생산되었고 활성은 대단히 높았다. 균주를 염료와 함께 10일간 배양했을 때 아조계 염료인 orange II, orange G, tropaeolin O 및 congo red의 탈색율은 각각 98.0%, 97.4%, 99.0% 및 95.3%로 나타났고 트리페닐메탄계 염료인 basic fuchsin, malachite green 및 crystal violet 들은 98.5%, 95.7% 및 99.4%로, 헤테로싸이클릭계 염료에 속하는 eosin Y, toludine blue, methyl blue 및 azur B는 각각 97.4% 98.7%, 99.9% 및 94.0%의 탈색율을 보였다. 송곳니구름버섯 BN2 균주에 의한 염료의 탈색은 주로 laccase에 의하여 이루어진다고 생각된다.

• 한국균학회지
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• 제22권4호
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• pp.325-331
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• 1994

좀구멍버섯균(Schizopora paradoxa)은 백색 부후 균으로 망간이 배지에 첨가되지 않으면 MNP와 GOX는 동시에 생성되지 않았으며 고농도의 망간 첨가시 (40ppm)에는 기본적 농도(11.15ppm)에 비해 MNP와 GOX가 모두 높은 활성을 보였다. 그러나 망간농도에 따른 균사생장량에는 별 차이가 없었다. 구리와 veratryl alcohol에 대해서도 같은 실험을 실시하였으나 농도에 따른 MNP와 GOX의 변화가 상관관계를 갖지 않는 것으로 나타났다. MNP와 GOX의 생성조절과 관련하여 cAMP의 영향을 조사한 결과 cAMP생성효소 억제제인 atropine을 농도별로 배양액에 첨가시 정도의 차이는 있었으나 MNP와 GOX생성이 동시에 억제되었다.

• 약학회지
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• 제43권5호
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• pp.598-605
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• 1999

The hot water and mathanol extracts of Artemisia argyi showed considerable cytotoxicities against H9(ATCC HTB 176) cancer cell with IC50 values of $48.6{\;}\mu\textrm{g}/ml$ and $51.9{\;}\mu\textrm{g}/ml$, respectively. These cytotoxicities were found to be dependent on the extract concentrations and culture days. CuZnSOD and MnSOD activities were significantly increased in the cytoplasm and mitochondria fractions of cancer cell, and media in the presence of Artemisia argyi. Such enhanced SOD activities were generally in the range of two to threefolds. In contrast to SOD, catalase and glutathione peroxidase activities were not detected at all. These results suggest that Artemisia argyi have generated $O_2^-$ in the mitochondria and cytoplasm of H9 cancer cell with concurrent induction of CuZnSOD and MnSOD in situ, which dismutate $O_2^-{\}to{\;}H_2O_2$. Without coordinated actions of catalase and/or glutathione peroxidase $H_2O_2$ is easily converted to very toxic OH and these reactive oxygen species together might have induced necrosis and/or apoptosis of H9 cell.

• Journal of the Korean Wood Science and Technology
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• 제32권6호
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• pp.14-22
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• 2004

The effect of cadmium ions on ligninolytic and decolourizing activities in cultures of two white-rot fungi, Cerrena unicolor and Trametes versicolor, were examined. Cadmium was added to the shallow stationary cultures growing on a liquid mineral medium. Both examined strains sorbed Cd ions in the first 24 hr of incubation. An appreciable stimulation of the activity of extracellular laccase (LAC) and inhibition of the extracellular manganese-dependent peroxidase (MnP) were simultaneously observed when 25 mgL^-1 and 50 mgL^-1 of cadmium ions were added to the cultures. On the other hand, the addition of cadmium ions also resulted in stimulating the decolorization activity of C. unicolor to decolorize Remazol Brilliant Blue R (RBBR) in the cultures, but decreasing it in the culture of T. versicolor, which is compared to the inhibition of MnP activity in this fungus. Our data indicate that the presence of Cd(II) ions can affect the ligninolytic activity of white-rot fungi. It was found that C. unicolor is a strain resistant to the presence of Cd ions in the liquid culture media, and has a potential to use this strain for bioremediation of sites contaminated with both heavy metals and aromatic pollutants.