• Journal of the Korean Wood Science and Technology
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• 제27권2호
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• pp.53-61
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• 1999

리그닌분해균 LSK-27 균주로부터 Manganese peroxidase (MnP) 생산을 위한 배지조건과 Mn(II) 첨가효과를 검토하였다. LSK-27균주에 의한 균체외 MnP 생산에는 탄소원보다 질소원의 영향이 크게 나타났다. 질소원으로는 peptone 이나 yeast extract와 같은 복합 유기질소원이 효과적이었으며 특히 질소원의 농도가 높은 조건에서 MnP activity가 우수하였다. 질소원으로서 peptone 농도 1.0%까지는 뚜렷한 activity 증가를 가져왔으나 1.5%이상의 농도에서는 MnP activity 증가효과가 크게 나타나지 않았다. 반면에 탄소원의 농도에 의한 MnP activity 증가효과는 glucose농도 1.0% 에서 3.0%까지 거의 비슷한 수준이었다. Mn(II)는 높은 MnP 유도효과를 나타냈으며 첨가농도 100ppm 까지 MnP activity의 증가효과를 보였고 그 이상의 농도에서는 MnP 생산이 억제되었다. Mn(II)는 배양 2일 후에 첨가하였을 때 MnP activitv 증가가 가장 높게 나타났다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.171-174
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• 2000

본 연구는 용존산소의 양에 따른 MnP의 생산양상을 체계적으로 연구하였으며, 과량의 산소 공급 시 MnP 생산이 저해될 수 있음을 산소공급에 따른 과량의 $H_2O_2$생산 측면에서 설명하고 있다. 또한 보다 높은 MnP 생산을 위한 산소공급방법을 제시하고자 하였다.

• Applied Biological Chemistry
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• 제47권1호
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• pp.22-26
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• 2004

리그닌 분해균인 느타리 버섯균(Pleurotus ostreatus) K-2946을 glucose-peptone-yeast(G-P-Y) 액체배지에서 배양 하여 망간퍼옥시데이즈를 생산하였다. 본 실험조건 하에서 리그닌 퍼옥시데이즈는 생산되지 않았다. 느타리버섯균 K-2946이 생산하는 망간 퍼옥시데이즈를 이온크로마토그래피, 겔크로마토그래피를 이용하여 정제하였다. 정제한 망간 퍼옥시데이즈의 분자량은 36400 Da이였고 pI는 3.95였다. 정제된 효소의 최적 pH는 5.0이었으며 최적온도는 $55^{\circ}C$이었다.

• 미생물학회지
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• 제45권1호
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• pp.82-85
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• 2009

내분비장애물질은 분해가 매우 어려워 자연계에서 먹이그물을 통하여 사람에게 농축 전달된다. 이들은 정상적인 내분비계에 혼란을 일으키며, 특히 성호르몬의 작용에 많은 피해를 준다. 이를 효율적으로 분해하고 이들의 에스트로겐 활성을 제거하고자 백색부후균의 하나인 아교버섯(Phlebia tremellosa)을 활용하여 4가지 내분비계 장애물질의 분해에 대한 실험을 수행하였다. 아교버섯의 manganese peroxidase (MnP) 활성을 높이기 위하여 구름버섯의 MnP 유전자를 아교버섯에 도입하여 형질전환체를 확보하였으며 이들은 유전적으로 MnP 활성을 안정되게 나타냈다. 내분비 장애물질을 분해하는 조건에서 내분비장애 물질에 따라 30${\sim}$45%의 분해율을 보인 야생형에 비하여 이 형질전환체들 중 T5는 70${\sim}$88%의 분해율을 보였으며 에스트로겐 활성의 제거에도 약 2배 향상된 능력을 보였다.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.130-134
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• 2003

A visual method for the selective screen Eng of lignin degrading enzymes, produced by white rot fungi (WRF), was investigated by the addition of coloring additives to solid media. Of the additives used in the enzyme production media, guaiacol and RBBR could be used for the detection of lignin peroxidase (LiP), manganese peroxidase (MnP) and lactase. Syringaldazine and Acid Red 264 were able for the detection of both the MnP and lactase, and the LiP and laccase, respectively, and a combination of these two additives was able to detect each of the ligninases produced by the WRF on solid media.

• 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.

• Journal of Microbiology and Biotechnology
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• 제32권2호
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• pp.248-255
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• 2022

Phlebia sp. MG-60 is the salt-tolerant, white-rot fungus which was isolated from a mangrove forest. This fungus expresses three kinds of manganese peroxidase (MGMnP) isozymes, MGMnP1, MGMnP2 and MGMnP3 in low nitrogen medium (LNM) or LNM containing NaCl. To date, there have been no reports on the biochemical salt-tolerance of these MnP isozymes due to the difficulty of purification. In present study, we established forced expression transformants of these three types of MnP isozymes. In addition, the fact that this fungus hardly produces native MnP in a high-nitrogen medium (HNM) was used to perform isozyme-selective expression and simple purification in HNM. The resulting MGMnPs showed high tolerance for NaCl compared with the MnP of Phanerochaete chrysosporium. It was worth noting that high concentration of NaCl (over 200 mM to 1200 mM) can enhance the activity of MGMnP1. Additionally, MGMnP1 showed relatively high thermo tolerance compared with other isozymes. MGMnPs may have evolved to adapt to chloride-rich environments, mangrove forest.

• Journal of Microbiology
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• 제43권6호
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• pp.503-509
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• 2005

The production of manganese peroxidase (MnP) by Irpex lacteus, purified to electrophoretic homogeneity by acetone precipitation, HiPrep Q and HiPrep Sephacryl S-200 chromatography, was shown to correlate with the decolorization of textile industry wastewater. The MnP was purified 11.0-fold, with an overall yield of $24.3\%$. The molecular mass of the native enzyme, as determined by gel filtration chromatography, was about 53 kDa. The enzyme was shown to have a molecular mass of 53.2 and 38.3 kDa on SDS-PAGE and MALDI-TOF mass spectrometry, respectively, and an isoelectric point of about 3.7. The enzyme was optimally active at pH 6.0 and between 30 and $40^{\circ}C$. The enzyme efficiently catalyzed the decolorization of various artificial dyes and oxidized Mn (II) to Mn (III) in the presence of $H_2O_2$. The absorption spectrum of the enzyme exhibited maxima at 407, 500, and 640 nm. The amino acid sequence of the three tryptic peptides was analyzed by ESI Q- TOF MS/MS spectrometry, and showed low similarity to those of the extracellular peroxidases of other white-rot basidiomycetes.

• 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.