• 제목/요약/키워드: Natural purification

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Purification and Characterization of Hrp1, a Homolog of Mouse CHD1 from the Fission Yeast Schizosaccharomyces pombe

  • Yong Hwan Jin;Eung Jae Yoo;Yeun Kyu Jang;Seung Hae Kim;Chee-Gun Lee;Rho Hyun Seong;Seung Hwan Hong;Sang Dai Park
    • Animal cells and systems
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    • 제2권4호
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    • pp.539-543
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    • 1998
  • Hrp1, of Schizosaccharomyces pombe, is a new member of the SW12/SNF2 protein family that contains a chromodomain and a DNA binding domain as well as ATPase/7 helicase domains. This configuration suggests that Hrp1 could be a homolog of mouse CHD1, which is thought to function in altering the chromatin structure to facilitate gene expression. To understand the enzymatic nature of Hrp1 we purified the 6-Histidine-tagged Hrp1 protein (6$\times$His-Hrp1) to homogeneity from a S. pombe Hrp1-overexpressing strain and hen examined its biochemical properties. We demonstrate that the purified 6$\times$His-Hrp1 protein exhibited a DNA-binding activity with a moderate preference to the (A+T)-rich tract in double-stranded NA via a minor groove interaction. However, we failed to detect any intrinsic DNA helicase activity from the purified Hrp1 like other SW12/SNF2 proteins. These observations suggest that the DNA binding activities of Hrp1 may be involved in the remodeling of the chromatin structure with DNA-dependent ATPase. We propose that Hrp1 may function in heterochromatins as other proteins with a chromo- or ATPase/helicase domain and play an important role in the determination of chromatin architecture.

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음나무 수피로부터 보체계 활성화 다당의 정제 및 특성 (Purification and Characterization of Complement System Activating Polysaccharide from the Bark of Kalopanax pictus N.)

  • 신금;라경수;백기현
    • Journal of the Korean Wood Science and Technology
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    • 제20권4호
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    • pp.73-84
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    • 1992
  • It was observed that the hot-water extract of the bark of Kalopanax pictus N. had the highest anti-complementary activity among the 11 kinds of forest materials. Methanol-and ethanol-soluble portions had low anti-complementary activities, but crude polysaccharide. HKP-0 had a high activity of 80%. HKP-0 contained 54.8% of total sugar and 27.9% of protein. The neutral sugars of HKP-0 consisted of mainly arabinose, galactose and glucose. HKP-4 fraction obtained by cetavlon treatment of HKP-0 showed the highest anti-complementary activity of 90%. The activity was not changed by pronase digestion bu decreased greatly by periodate oxidation. HKP-4 consisted of mainly arabinose and glucose with molar ratio of 1.0 : 22.4, HKP-4-I, an unabsorbed fraction from HKP-4 on DEAE Sepharose CL-6B column showed higher yield and activity than those of absorbed fractions. HKP-4-I was homogeneous, and its molecular weight was about 25,000. HKP-4-I contained 84.0% of neutral sugar and consisted of arabinose and glucose with molar ratio of 1.0 : 11.2. The anti-complementary activity of HKP-4-I was not decreased by the treatment of polymyxin B, and the polysaccharide activated both classical and alternative pathway in complement system. Void volume fraction obtained from HKP-4-I hydrolyzed with ${\alpha}$-amylase on Sephadex G-25 column only had a high anti-complementary activity.

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하천에 있어서 자연성의 보전, 정비, 창출에 관한 연구 II - 원성천과 풍서천의 하천식생구조를 대상으로 - (A Study on the Conservation Rehabilitation and Creation of Naturalilty of Rivers - River Vegetation Structure of Wonsungcheon and Pungseocheon)

  • 방광자;이진희;설종호;강현경;박성은
    • 환경영향평가
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    • 제7권2호
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    • pp.53-64
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    • 1998
  • This study was performed to build up the ecological guidelines to grasp the structure of the vegetation change which is due to river rehabilitation. Anyway, river ecosystem and function has been destroyed owing to river development. It is important that river vegetation supplies ecological corridor and biotope. Two survey sites(Wonsungcheon and Pungseocheon)were investigated in the aspect of plant ecosystem and structure to settle the practical concept of river ecosystem. Each survey site was subdivided to five plots. The site was surveyed through the belttransect method. Wonsungcheon gets more seriously polluted as it runs to the urban area. In other words, there are On the other hand, Pungseocheon has more naturality but its downstream is under the pressure of various wood plants in the upstream area, but downstream area is dominated by naturalized plants such as Bidens frondosa, Panicum dichotomiflorum, etc. Riverbank of downstream has been changed into farm and parking lot. development. It should be preserved definitely because it still has abundant naturality and wetland which formed a biotope. The objective of the research is to find out the river retrogression and maintenance methods based on the riparian vegetation structure. To manage the river ecologically, hydrophytes should be induced partly for natural purification after the riverside is rehabilitated. The vegetation should be induced step by step to restore natural river and steady monitoring and research are required.

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생물다양성 증진을 위한 유휴농경지의 습지 복원 적지선정에 관한 연구 - 군포시 반월천 유역 사례로 - (Wetland Restoration Site Selection for Promoting Biodiversity in Abandoned Rice Paddy Fields - Focusing on Gounpo Ban-wol Stream Watershed -)

  • 임유라;김귀곤
    • 한국환경복원기술학회지
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    • 제12권1호
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    • pp.52-66
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    • 2009
  • The increase of rice productivity, the decrease of rice demand, aging farmers, and the market-opening of rice along with the domestic and international conditions changes have been led to the increase of the abandoned rice paddy fields. Such abandoned rice paddy fields have been left in many areas and most of them have become wetlands. The purpose of this study is methodology of selection to the wetland for development in the abandoned rice paddy fields along with surroundings. The selection of habitat suitability index assessment study to conservation and restoration was done through hydrology, wild animals habitation, wetland vegetation, outside disturbance, and natural purification capacity assessment. In addition, the same method was applied to assessment items and standards for both the restoration site selection and the type of restoration. Both assessment items and standards were applied to the sites, drawn on the maps, and overlayer for the comprehensive map, and then the wetland suitability index was applied to the suitable site. The development technique was applied to 10 sites near the Banwol Stream watershed in Gunpo, Gyeonggi province. The selection of conservation, restoration, and the optimal sites can lead to not only the biodiveristy increase in agricultural or semi natural areas but also to the establishment of ecological networks in national level.

월계화 추출물의 면역억제 효능 연구 (Study on Immunosuppressive Effects of Rosa Chinensis Jacq. Extract)

  • 김경신;박재원;채순기;강정수;김병수
    • 동의생리병리학회지
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    • 제25권3호
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    • pp.459-465
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    • 2011
  • The nuclear factor of activated T cells (NFAT) protein induces transcriptions of cytokine genes including IL-2 for T-cell activation. Normally activation of NFAT is important to induce immune responses but excessive NFAT activation provokes immunopathological reactions such as autoimmunity, transplant rejection, and inflammation. Thus, for the treatment of autoimmune diseases drugs repressing the activation of NFAT have been searched. In this study, immnunosuppressive effects of Rosa chinensis Jacq. extracts identified as a potent NFAT inhibitor from a natural product library were examined. NFAT reporter assay, MTS assay, real time PCR, IL-2 ELISA, MLR, and FACS (Fluorescent Activated Cell Sorting) were used to measure inhibitory immunocyte activities of Rosa chinensis Jacq. The variety of natural products have been screened and some were found to show inhibitory activities against the NFAT transcription factor. Among them, extract of Rosa chinensis Jacq. showed an strong inhibitory effect on the activation of NFAT without affecting cell viability. Levels of IL-2 transcripts as well as IL-2 protein were decreased with treatment of Rosa chinensis Jacq. extract. In addition, immunosuppressive activity of Rosa chinensis Jacq. extract was exhibited in the mixed leukocytes reaction. The increasement of CD4+CD25+ (Treg) immunocyte was also detected in the analysis using FACS after applying Rosa chinensis Jacq. extract. Immunosuppressive effects of the Rosa chinensis Jacq. extracts were clearly demonstrated in the present study. In addition, Rosa chinensis Jacq. extract also positively affected regulatory T cell induction. Further investigations in particular on purification of single substance responsible for the immunosuppressive effects from the extract and analysis on possible actions of the extract in interfering cell signaling and cytokine production will be needed.

Phenolic Constituents from the Flowers of Hamamelis japonica Sieb. et Zucc.

  • Yim, Soon-Ho;Lee, Young Ju;Park, Ki Deok;Lee, Ik-Soo;Shin, Boo Ahn;Jung, Da-Woon;Williams, Darren R.;Kim, Hyun Jung
    • Natural Product Sciences
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    • 제21권3호
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    • pp.162-169
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    • 2015
  • Hamamelis japonica (Hamamelidaceae), widely known as Japanese witch hazel, is a deciduous flowering shrub that produces compact clumps of yellow or orange-red flowers with long and thin petals. As a part of our ongoing search for phenolic constituents from this plant, eleven phenolic constituents including six flavonol glycosides, a chalcone glycoside, two coumaroyl flavonol glycosides and two galloylated compounds were isolated from the flowers. Their structures were elucidated as methyl gallate (1), myricitrin (2), hyperoside (3), isoquercitrin (4), quercitrin (5), spiraeoside (6), kaempferol 4'-O-β-glucopyranoside (7), chalcononaringenin 2'-O-β-glucopyranoside (8), trans-tiliroside (9), cis-tiliroside (10), and pentagalloyl-O-β-D-glucose (11), respectively. These structures of the compounds were identified on the basis of spectroscopic studies including the on-line LCNMR-MS and conventional NMR techniques. Particularly, directly coupled LC-NMR-MS afforded sufficient structural information rapidly to identify three flavonol glycosides (2 - 4) with the same molecular weight in an extract of Hamamelis japonica flowers without laborious fractionation and purification step. Cytotoxic effects of all the isolated phenolic compounds were evaluated on HCT116 human colon cancer cells, and pentagalloyl-O-β-D-glucose (11) was found to be significantly potent in inhibiting cancer cell growth.

Characterization of Two Forms of Glucoamylase from Traditional Korean Nuruk Fungi, Aspergillus coreanus NR 15-1

  • HAN YOUNG JIN;YU TAE SHICK
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.239-246
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    • 2005
  • Some characteristics of two forms of glucoamylase (glucan 1 A-$\alpha$-glucosidase, EC 3. 2. I. 3) purified from Aspergillus coreanus NR 15-1 were investigated. The enzymes were produced on a solid, uncooked wheat bran medium of A. coreanus NR 15-1 isolated from traditional Korean Nuruk. Two forms of glucoamylase, GA-I and GA-II, were purified to homogenity after 5.8-fold and 9.6-fold purification, respectively, judged by disc- and SDS-polyacrylamide gel electrophoresis. The molecular mass of GA-I and GA-II were estimated to be 62 kDa and 90 kDa by Sephadex G-1OO gel filtration, and 64 kDa and 91 kDa by SDS-polyacrylarnide gel electrophoresis, respectively. The optimum temperatures of GA-I and GA-II were 60$^circ$C and 65$^circ$C, respectively, and the optimum pH was 4.0. The activation energy (Ea value) of GA-I and GA-II was 11.66 kcal/mol and 12.09 kcal/mol, respectively, and the apparent Michaelis constants (K_{m}) of GA-I and GA-II for soluble starch were found to be 3.57 mg/ml and 6.25 mg/ml, respectively. Both enzymes were activated by 1 mM Mn^{2+} and Cu^{2+}, but were completely inhibited by 1 mM N­bromosuccinimide. The GA-II was weakly inhibited by 1 mM p-CMB, dithiothreitol, EDTA, and pyridoxal 5-phosphate, but GA-I was not inhibited by those compounds. Both enzymes had significant ability to digest raw wheat starch and raw rice starch, and hydrolysis rates of raw wheat starch by GA-I and GA-II were 7.8- and 7.3-fold higher than with soluble starch, respectively.

연못을 이용한 동절기 인공습지 오수처리수의 추가 처리 (Pond System for Further Polishing of Constructed Wetland Effluent during Winter Season)

  • 윤춘경;전지홍;김민희;함종화
    • 한국농공학회지
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    • 제44권4호
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    • pp.139-148
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    • 2002
  • Pilot study was performed to examine the feasibility of the pond system for further polishing of treatment wetland effluent from December 2000 to June 2001. The wetland system used for the experiment was highly effective to treat the sewage during the growing season, but it was less effective and its effluent was still high to discharge to the receiving water body. Therefore, the wetland effluent may need further treatment to prevent water quality degradation. Pond system could be used to hold and further polish the wetland effluent during the winter season and ots feasibility was evaluated in this study. Additional water quality improvement was apparent in the pond system during winter season, and the pond effluent could be good enough to meet the effluent water quality standards if it is properly managed. Timing of the pond effluent discharge appears to be critical for pond system management because it is a closed system and whole water quality constituents are affected by physical, chemical, and biological pond environments. Once algae started to grow in mid-April, constituents in the pond water column interact each other actively and its control becomes more complicated. Therefore, upper layer of the pond water column which is clearer than the lower layer my need be discharged in March right after ice cover melted. In the experiment, water quality of the upper water column was markedly clear in March than ant other times probably because of freezing-thawing effect. The remaining lower water column could be further treated by natural purification as temperature goes up or diluted with better quality of wetland effluent for appropriate water uses. This study demonstrated the feasibility of pond system for subsequent management of wetland effluent during the winter season, however, more study is needed for field application.

Ergostane-Type Steroids from Korean Wild Mushroom Xerula furfuracea that Control Adipocyte and Osteoblast Differentiation

  • Lee, Seoung Rak;Choi, Jin Hee;Ryoo, Rhim;Kim, Jin-Chul;Pang, Changhyun;Kim, Seon-Hee;Kim, Ki Hyun
    • Journal of Microbiology and Biotechnology
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    • 제30권11호
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    • pp.1769-1776
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    • 2020
  • As part of our current work to discover structurally and/or biologically novel compounds from Korean wild mushrooms, we isolated five ergostane-type steroids (1-5) from the fruiting bodies of Xerula furfuracea via repeated column chromatographic separations and HPLC purification. The chemical structures of the isolated steroids were shown to be (22E,24R)-24-methylcholesta-4,22-diene-3,6-dione (1), ergosta-7,22-diene-3β,5α,6β-triol (2), ergosta-7,22-diene-3β,5α,6β,9α-tetraol (3), (22E,24R)-5α,8α-epidioxyergosta-6,22-diene-3β-ol-3-O-β-D-glucopyranoside (4), and (22E,24R)-5α,8α-epidioxyergosta-6,9,22-triene-3β-ol-3-O-β-D-glucopyranoside (5)based on comparison of the data regarding their spectroscopic and physical properties with those of previous studies. Notably, this is the first report on the presence of the identified steroids (1-5) in this mushroom. We tested compounds 1-5 to determine their effects on adipogenesis and osteogenesis in the mouse mesenchymal stem cell line C3H10T1/2 and found that compounds 4 and 5 suppressed the differentiation of stem cells into adipocytes. Notably, in addition to its suppressive effect on adipogenesis, compound 5 was also shown to promote the osteogenic differentiation of stem cells. These findings demonstrate that the bioactive compounds isolated might be effective for the treatment of menopause-associated syndromes, such as osteoporosis and obesity, as the isolated compounds were shown to suppress adipogenesis and/or promote osteogenesis of stem cells.

고염 원시박테리아(Halococcus agglomeratus)에 존재하는 TPx 분리 및 생화학적 특성연구 (Purification and characterization of TPx from archeabacteria, Halococcus agglomeratus)

  • 최용수;차미경;김일한
    • 자연과학논문집
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    • 제14권2호
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    • pp.67-82
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    • 2004
  • 고염에서 자라는 원시 박테리아인 Halococcus agglomeratus에서 Thiol-specific antioxidant 활성을 보이는 분자량이 22-kDa인 향산화 단백질을 순수 분리 정제하여 향산화 활성의 특성을 조사하였다. 그 결과 진핵 세포의 Thiol-specific antioxidant protein (TSA of TPx)과 유사한 활성을 갖는 것을 확인 할 수 있었다. 정제된 Thiol-specific antioxidant protein 은 환원제로 thiol 성분을 갖는 비효소적 금속 촉매 산화계( $Fe^{3+}$, $O^2$, DTT 또는 2-mercatoethanol : thiol- MCO system)에 의하여 Glutamine Synthetase (GS)의 불활성화를 방어하고 Ascorbate 같은 nonthiol 성환원제를 갖는 금속 촉매 산화계 ( $Fe^{3+}$, $O^2$, Ascorbatenol: nonthiol- MCO system)에 의해서는 GS의 불활성화를 방어하지 못하였다. 이것은 환원형 thiol성분이 항산화 단백질의 항산화 활성에 전자 공여체로 요구되어 지기 때문이라고 판단된다. 이 단백질은 다른 TPx와는 다르게 100%의 활성을 나타내려면 NaCl의 농도가 500mM이상이 되어야 한다. 이상의 결과는 원시 박테리아에도 TPx가 존재하여 활성 산소종을 제거하는 생화학적 역할을 수행함을 시사하고 있다.

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