• Title/Summary/Keyword: Natural purification

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Lymphocytes Mitogenic and Immunochemical Characteristics of the Immunomodulating Lectins, MLA, from Marine Natural Products (해양 천연물에서 분리한 면역기능 조정제 렉틴 MLA의 림프구 자극분열효과 및 면역화학적 특성)

  • 전경희;김장환;정시련
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.252-259
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    • 1995
  • Isolation, purification and characterization of biophysicochemical properties of the three new lectins, MLA-I, MLA-II, MLA-III from the hemolymph of Meretrix lusoria have been reported previously. A series of immunochemical studies were investigated in this work. The three lectins were revealed as having partial identity each other by immunodiffusim and immunoelectrophoresis. These results suggest that MLA lectins are isolectins having similar biophysicochemical properties. Particularly, MLA-I proved to be a potent mitogen for murine splenic as well as human peripheral lymphocytes, and the optimum mitogenic dose were 62.5 and 1.95 $\mu\textrm{g}$/ml, respectively.

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An Experimental Study on the Properties of Hwangtoh Mortar for the Application of Construction Material (건축적 적용을 위한 황토모르터의 물성에 관한 실험적 연구)

  • Lee, Hyun-Chul;Lee, Gun;Go, Seong-Seok
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2006.11a
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    • pp.43-46
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    • 2006
  • The Hwangtoh is one of the traditional construction material used in wall, plastering material, and ondol (Korean underfloor heating system) with stone and wood. It is an important greenness material and it has much advantages such as; high storage of heat, auto-purification, antibiotic ability, and emission of far infrared rays. But, it is not developed and not used in modern construction because of its low strength and properties of dry shrinkage crack. According to the recent researches and studies, it is evaluated for natural pozzolanic material like flyash or pozzolan. It's possibility on construction material is high because it's chemical and mineralogical proportion is like as Metakaolin and Kaolinite. In this point of view, this study aims to analyze the physical properties on Hwangtoh mortar through an experiment with various activation condition of Hwangtoh, which is natural pozzolanic material, for the purpose of increase the using possibility in construction material.

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칠성장어 (Lampetra japonica)젖 산수소이 탈효소 동위효소의 정제 및 면역화학

  • Jo, Seong-Gyu;Park, Seon-Yeong;Yeom, Jeong-Ju
    • The Korean Journal of Zoology
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    • v.36 no.4
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    • pp.505-513
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    • 1993
  • 칠성장어(Lampetro juponico) 골격근의 젖산수소이탈효소(EC 1. 1. 1.27, Lactate dehvdrogenase LDH) 동위효소 꺽지(Coreoperco herzi)의 LDH Aa 동위효소 및 소(Bos taurus)의 LDH B4 동위효소를 affinity chromatography로 정제하였고, 대구(Gadus macrocephcfus)의 LDH Ca 동위효소를 affinity chromatography와 OEAE-Sephacel chromatography로 정제하였다. 칠성장어 골격근 LDH 동위효소는 affinity chromatography에서 buffer를 유입한 후 용출되는 분획에서 모두 확인되었고, 정제한 칠성장어 골격근의 LDH에 대한 항체는 꺽지 LDH A4, C4 및 54 동위효소, 대구 LDH A4 및 B4 동위효소 그리고 생쥐(MUS musculus) LDH A4 등위효소와 복합체를 형성하였다. 칠성장어 골격근조직에서는 비각우 및 Ldh절가 발현되고 하부단위체 A의 구조는 하부단위체 B의 구조와 유사하며, 하부단위체 A는 진화상 보존적 이지만 하부단위체 C는 진화속도가 상당히 빠른 것으로 사료된다.

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Studies on the Constituents of Rhamnella frangulioides (까마귀베개뿌리의 성분에 관한 연구)

  • Yoo, Seung-Jo;Song, Kyung-Shik;Kwak, Jong-Hwan;Han, Byung-Hoon;Ryu, Jae-Ha
    • Korean Journal of Pharmacognosy
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    • v.20 no.3
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    • pp.147-148
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    • 1989
  • From the root of Rhamnella frangulioides(=Microrhamnus frangulioides), three compounds (Comp. I ; $mp\;140{\sim}142^{\circ},\;Comp.\;II\;;\;mp\;196^{\circ},\;Comp.\;III\;;\;mp\;136{\sim}138^{\circ})$ were isolated by silica gel column chromatographic purification which were identified as ${\beta}-sitosterol$, chrysophanol, 1-methyl-2-carboxymethyl-3-methoxy-4,8-dihydroxy andthraquinone, respectively by spectral analysis.

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High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

Purification and Identification of a Natural Antioxidant Protein from Fertilized Eggs

  • Yang, Shaohua;Wang, Lulu;Wang, Ying;Ou, Xiaoqian;Shi, Zhaoyuan;Lu, Chongchong;Wang, Wei;Liu, Guoqing
    • Food Science of Animal Resources
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    • v.37 no.5
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    • pp.764-772
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    • 2017
  • Fertilized hen eggs are rich in a variety of bioactive ingredients. In this study, we aimed to obtain an antioxidant protein from fertilized eggs and the radical scavenging abilities on 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (${\bullet}OH$), superoxide anion ($O^{2-}{\bullet}$) were used to evaluate the antioxidant activity of the purified protein. During 20 d of incubation, the radical scavenging ability of protein extracted from fertilized eggs exhibited significantly differences and the protein on day 16 showed higher antioxidant capacity. Based on this, the antioxidant protein of the samples on day 16 were isolated for the follow-up study. With a molecular weight 43.22 kDa, the antioxidant protein was purified by Diethylaminoethyl cellulose -52 (DEAE-52) column and Sephadex G-100. The LC-MS analysis showed that the purified protein molecular weight was 43.22 kDa, named D2-S. The sequence of amino acids was highly similar to ovalbumin and the coverage reached to 84%. The purified protein showed a radical scavenging rate of $52.34{\pm}3.27%$ on DPPH and $63.49{\pm}0.25%$ on ${\bullet}OH$, respectively. Furthermore, the C-terminal amino acid sequence was NAVLFFGRCVSP, which was consistent with the sequence of ovabumin. These results here indicated that purified protein may be a potential resource as a natural antioxidant.

Production of Oleamide, a Functional Lipid, by Streptomyces sp. KK90378

  • Kwon, Jeong-Ho;Hwang, Sung-Eun;Han, Jae-Taek;Kim, Chang-Jin;Rho, Jung-Rae;Shin, Jong-Eon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1018-1023
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    • 2001
  • Oleamide (cis-9-octadecenamide) is endogenous primary amide of fatty acid that is produced in small amounts in animal brains. It is known to induce sleep and to lower temperature by destroying the lipid plasma membrane structure of cells, thereby disclosing gap junction channels. To develop a new biological production method for oleamide, a screening program was conducted to isolate a microorganism producing oleamide. Among 1,500 soil microorganisms tested, KK90378 exhibited a potent positive reaction with Dragendoff`s reagent, used to detect the primary amide of oleamide. KK90378 was identified as a Streptomyces species based on cultural and morpohological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extrat. Streptomyces sp. KK90378 produced oleamide 3 days after culture at $28^{\circ}C$, pH 7.2 A series of purification steps, including hexane extraction, silica gel column, and preparative thin layer chromatographies, were performed for the purification of oleamide. A spectrophotometric analysis using $^1H$, $^13C$-NMR, and GC-MS confirmed that the chemical structure of the purified oleamide was identical to that of authentic oleamide.

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Purification and Characterization of Heparin Lyase I from Bacteroides stercoris HJ-15

  • Kim, Wan-Seok;Kim, Byung-Taek;Kim, Dong-Hyun;Kim, Yeong-Shik
    • BMB Reports
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    • v.37 no.6
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    • pp.684-690
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    • 2004
  • Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

Development of Constructed Wetland using Sand and Oyster shell for Sewage Treatment (모래와 굴패각을 이용한 인공습지 오수처리장치 개발)

  • Park, Hyun-Geoun;Lee, Chun-Sik;Lee, Hong-Jae;Seo, Dong-Cheol;Heo, Jong-Soo
    • Journal of Korean Society on Water Environment
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    • v.20 no.5
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    • pp.437-446
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    • 2004
  • To develop sewage treatment apparatus by natural purification method, the sewage treatment plant that consisted of aerobic and anaerobic plot was constructed. And then, the effects of treatment conditions on the removal of pollutants in the relation to sewage loading, sewage injection method and season according to the pebble kind. Removal rate of BOD and COD according to the sewage loading in effluent were over 95 and 77%, respectively. Removal rate of nitrogen in treated water by aerobic plot and effluent using sand were about 22~40 and 49~75%, respectively. Those of effluent using sand 75%+ oyster shell 25% and sand 50%+ oyster shell 50% as filter media in comparison with using sand were about 7~25 and 16~23%, respectively. Removal rate of phosphorus in treated water by aerobic plot and effluent using sand as filter media were about 30~36 and 52~56%, respectively. Those of effluent using sand 75%+ oyster shell 25% and sand 50%+ oyster shell 50% in comparison with using sand as filter media were about 11~40 and 12~45%, respectively. Removal efficiency of BOD and COD according to the intermittent injection method of sewage were slightly decreased, but those of nitrogen and phosphorus were little varied in comparison with continuous injection method. Removal efficiency of BOD and COD in winter in comparison with the others were little varied, but those of nitrogen and phosphorus slightly decreased.

Purification and Characterization of an Antioxidant Protein from Fertilized Eggs

  • Yang, Shaohua;Tang, Zhengjiang;Tang, ShanShan;Zhang, Tingfang;Tang, Fei;Wu, Yu;Wang, Ying;Wang, Lu Lu;Liu, Guoqing
    • Food Science of Animal Resources
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    • v.36 no.6
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    • pp.791-798
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    • 2016
  • Free radicals may attack cells or tissue, leading to chronic diseases, and antioxidant consumption is potentially useful for removing free radicals. Egg proteins may be used as potential sources of antioxidant considering their ability of scavenging free radicals to apply for food or cosmetics industry. In this study, we obtained a natural antioxidant protein from fertilized eggs, which was a dietary supplement in some Asian countries. Meanwhile, antioxidant activities of these proteins were evaluated using different oxidation systems. With increasing incubation time, the antioxidant activity of these proteins increased during 15 d of incubation. The samples on day 15 were performed for isolation of antioxidant protein. The protein, named P4-1 (MW, 45 kDa), was isolated and purified by consecutive chromatographic methods. P4-1 contained 17 amino acids, which was determined by liquid chromatography-mass spectrometry and Amino Acid Analyzer. Moreover, the amino acid sequence was highly similar to that of ovalbumin. Differential scanning calorimetry showed that the denaturation temperature of P4-1 was $57.16^{\circ}C$. Furthermore, P4-1 suggested high oxygen radical-absorbance activity in ${\cdot}OH$ assays, and its antioxidant activity was stable at $30-50^{\circ}C$ in acidic and neutral pH. Thus, these results revealed that P4-1 may be a potential resource as a natural antioxidant.