• 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.

• Biomolecules & Therapeutics
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• 제12권3호
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• pp.189-193
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• 2004

Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

• 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.

• 생명과학회지
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• 제12권4호
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• pp.392-398
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• 2002

Lignocellulose 분해 이용을 위해 토양 등에서 lignin분해 능력이 우수한 세균을 분리 및 동정함으로서 방향족 화합물의 이용을 위한 생물전환의 기초를 확보하고자하였다. 토양 등의 시료로부터 38주의 리그닌 분해 균주를 분리하였다. 분리된 LG2를 공시 균주로 선정하여 형태학적, 배양학적 및 생리학적 특성을 조사한 결과 Pseudomonas sp. LG2로 명명하였다. 이 균주는 리그닌을 분해 할 수 있었으며, 리그닌 함유 배지에 배양하여 배양 분해 산물을 HPLC로 분석한 결과 다수의 방향족 화합물을 생산하였다 Poylacrylamide gel활성 분석에 의해서 3종류로 구성된 peroxidase가 조사되었다.

• Journal of Microbiology
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• 제43권6호
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• pp.569-571
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• 2005

White-rot fungi have the following enzyme systems for lignin degradation: laccase, lignin peroxidase and manganese peroxidase. There are other types of peroxidases related to lignin degradation, one of which we have cloned a cDNA gene of manganese-repressed peroxidase (MrP) in Trametes versicolor isolated in South Korea. The mrp transcript level has been decreased by $1{\mu}M\;of\;Mn^{2+}$.

• KSBB Journal
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• 제32권2호
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• pp.96-102
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• 2017

The lignocellulose that is a major component of spent coffee ground was degraded and saccharified. To implement the spent coffee, after several pre-treatments, inoculation of Phanerochaete chrysosporium and solid-state fermentation were conducted. The optimal temperature of the enzymes (lignin peroxidase, manganese peroxidase, xylanase, laccase, and cellulase) for degradation of lignocellulose by P. chrysosporium was found. We also measured the maximum activity of enzymes (lignin peroxidase 0.15 IU/mL, manganese peroxidase 0.90 IU/mL, laccase 0.11 IU/mL, cellulase 5.87 IU/mL, carboxymethyl cellulase 9.52 IU/mL, xylanase 1.16 IU/mL) used for the process. As a result, 4.73 mg/mL of reduced sugar was obtained and 61.02% of lignin was degraded by solid state fermentation of P. chrysosporium on spent coffee ground.

• 미생물학회지
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• 제36권3호
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• pp.228-235
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• 2000

지금까지의 Phanerochaete chrysosporium을 이용한 Lignin Peroxidase(LiP) 의 생산은 최적화 되지 못한 배양 조건으로 인해 매우 낮은 활성만을 얻을수 있었으며. 그 배양조건 또한 100% 산소의 주입을 요하는 일반적으로는 재현하기 어려운 조건이었다. 본 연구에서는 Phanerochaete chrysosporium PSBL-1 균줄르 이용, 다양한 배양조건의 변화를 통해 LiP의 과량생산과 그에 따른 isozyme 조성의 변화를 확인하고자 하였다. 최종적으로 $Mn^{2+}$를 제거한 배지에 질소 원인 diammonium을 48mM 로 첨가하고, 안정제로서 veratryI alcohol을 2 mM로 첨가하여, sponge에 의한 고착배양을 실시한 결과 1,800 units/l의 LiP가 생산되었다. 이러한 활성의 증가는 기존에 보고된 PSBL-1 균주의 LiP 최대 생산량인 700units/l와 비교 , 약 2.57배의 생산량 증대를 나타낸다. 생산된 LiP isozyme 분석결과 $Mn^{2+}$(2.73 mM)와 diammonium(48 mM)을 첨가하였을 경우 H1 isozyme의 과량 생산이 확인되었다. 생산된 LiP(0.4 units)를 이용하여 각각의 아조계 염료(acid yellow 9, congo red, orange II; each 50$\mu$M)에 대한 탈생능을 확인한 결과, 2분 이내에 이들 염료에 대한 탈색이 이루어지는 것을 확인할 수 있었고 분해 산물이 축적되는 것을 확인할 수 있었다.

• 대한임상검사과학회지
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• 제38권2호
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• pp.87-93
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• 2006

This study was done to know a kind and change (transition) of enzymes produceed by Streptomyces halstedii ssp. scabies SA1-27 and Streptomyces violaceusinger C1-6 which showed good lignolytic activity and a good decolorization ratio of remazol brilliant blue R(RBBR) dye. These strains were isolated from soil and identified by the author. The basal medium containg 0.2% glucose was used to measure enzyme activity, Lignin peroxidase 1 (Lip 1) was measured by the methods of Choi, and Bourbonnais and Paice. Lignin peroxidase 2 (Lip 2) was measured by the methods of Ishida et al and Ramachandra et al using 2.4-dichlorophenol(2.4 DCP), manganese peroxidase(Mnp), veratryl alcohol oxidase (VAO), and laccase. They were measured by each of the methods of Choi and Paszczynski et al, and Bourbonnais and Paice, and De Jong et al. In the results, the kind of enzymes produced by Streptomyces halstedii ssp. scabies SA1-27 were Lip 1, Lip 2, VAO, and laccase, and their activities indicated the highest value as each 4.95 nmol/mg protein, $8.45({\times}100^{-3})unit$, 10.25 nmol/mg protein, 9.20 nmol/mg protein on the sixth day of the culture and decreased gradually over time. The kind of enzymes produced by Streptomyces violaceusinger C1-6 were Lip 1, Lip 2, Mnp, VAO, and laccase, and their activities indicated the highest value as each 4.90 nmol/mg protein, $13.85({\times}100^{-3})unit$, 3.10 nmol/mg protein, 11.30 nmol/mg protein, 4.45 nmol/mg protein on the sixth day of the culture and decreased gradually over time. Consequently, the author knew the fact that there were few differences in the kind and quantity of enzymes produced by the two Streptomyces strains, but all enzyme activities indicated the highest value on the sixth day of the culture and decreased gradually over time.

• 한국미생물·생명공학회지
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• 제30권3호
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• pp.287-292
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• 2002

본 연구에서는 경안천 유역에 있는 주유소 오염토양으로부터 대표적인 아조계 염료 congo-red분해능이 우수한 토양 방선균, SMA-2를 분리 선별하고, SMA-2의 배양조건을 최적화하여 2,4-dichlorophenol의 산화에 관여하는 actinomycetes lignin peroxidase(ALiP)의 활성도에 대한 특성을 규명하였다. 기존에 보고된 방선균 유래 ALiP와는 달리, SMA-2유래 ALiP효소는 pH6에서 최고의 활성을 보였으며, 2,4-DCP와 $H_2$$O_2$에 대한 Km값이 각각 0.21mM과 8.5mM로 측정되었다. 또한 최적화된 배지에서 성장한 SMA-2의 배양액은 2,4-dichlorophenol 뿐만 아니라 2-chlorophenol, 2,4,6-trichlorophenol, 2,6-dichlorophenol, phenol, 4-chloropheno떼 대해서도 산화능을 보임으로써, 토양 방선균을 이용한 염화 페놀계 화합물이 포함된 오염 토양의 생복원 가능성을 제시하였다.

• Journal of the Korean Wood Science and Technology
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• 제28권4호
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• pp.17-24
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• 2000

The peroxidase activity was localized cytochemically to get an insight into its precise function in lignin biosynthesis. In this work, cerium chloride ($CeCl_3$) was used as a trapping agent for hydrogen peroxide ($H_2O_2$) generated from peroxidase. Seasonal variation of peroxidase activities in cambial region of Populus, Pinus, and Ginkgo was investigated at subcellular levels. Under transmission electron microscopy, electron dense deposits of cerium perhydroxide formed by reaction with $H_2O_2$ were observed in cambium and its immediate derivatives. The staining with $CeCl_3$ in cambium varied with growth seasons. The strongest $H_2O_2$ accumulation, regardless of tree species, appeared in May. Staining pattern of $CeCl_3$ in the cambium of poplar indicated that the production of peroxidase started in March before the opening of buds and reached the highest in May and then declined in August. Ginkgo and Pinus showed relatively late generation of $H_2O_2$ production when compared with Populus. Although Ginkgo and Pinus are classified into gymnosperms, however, the generation of peroxidase production and its duration was different from each other. Little staining appeared in all the tree samples collected in September before falling the leaves.