• 제목/요약/키워드: Membrane chromatography

검색결과 261건 처리시간 0.03초

Isolation of a Calcium-binding Peptide from Chlorella Protein Hydrolysates

  • Jeon, So-Jeong;Lee, Ji-Hye;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • 제15권4호
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    • pp.282-286
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    • 2010
  • To isolate a calcium-binding peptide from chlorella protein hydrolysates, chlorella protein was extracted and hydrolyzed using Flavourzyme, a commercial protease. The degree of hydrolysis and calcium-binding capacity were determined using trinitrobenzenesulfonic acid and orthophenanthroline methods, respectively. The enzymatic hydrolysis of chlorella protein for 6 hr was sufficient for the preparation of chlorella protein hydrolysates. The hydrolysates of chlorella protein were then ultra-filtered under 5 kDa as molecular weight. The membrane-filtered solution was fractionated using ion exchange, reverse phase, normal phase chromatography, and fast protein liquid chromatography to identify a calcium-binding peptide. The purified calcium-binding peptide had a calcium binding activity of 0.166 mM and was determined to be 700.48 Da as molecular weight, and partially identified as a peptide containing Asn-Ser-Gly-Cys based on liquid chromatography/electrospray ionization tandem mass spectrum.

대두 사포닌의 활성에 관한 기전 연구

  • 성미경
    • Journal of Nutrition and Health
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    • 제28권10호
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    • pp.1022-1030
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    • 1995
  • Saponins are glycosidic compounds present in many plant foods. They are characterized by their ability to lyse cell membranes due to their surface-active properties. Saponins are believed to interact primarily with cholesterol in the cell membrane. In this study, the interaction of soybean(SS) with cell membrane was investigated using erythrocytes as a model. Mechanisms of interaction was also investigated by measuring their binding capacity with different membrane lipid fractions. Throughout the study, gypsophilla saponin(GS) and quillaja saponin(QS) were used to evaluate the membranolytic activity of soybean saponins. All saponins released hemoglobin in a concentration-dependent manner. SS induced 40% hemolysis at the concentration of 400 ppm, however there was no increase in hemoglobin release above 400ppm concentration. 5ppm of GS and 8 ppm of QS hemolyzed 100% of erythrocytes. Isolation of SS fractions by thin layer chromatography revealed that only one non-polar saponin possesses strong hemolytic activity. When saponins were incubated decreased the release of cholesterol. When the hemolytic activity of saponins was measured in the presence of other major membrane lipid components, sphingomyelin significantly reduced the hemolytic activity of SS, while cholesterol reduced the activity of QS. GS showed high affinity to other component(s) in the incubation media as well as lipids. These results suggest that the membranolytic activity of saponins are related to their specific chemical structure, which determines the interaction behavior between saponins and different membrane components, and thereby influence the biological activity.

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고분자막을 이용한 효소의 분리 (Separation of enzymen with polymeric membrane)

  • 이제권;정용섭;홍석인
    • 한국막학회:학술대회논문집
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    • 한국막학회 1991년도 추계 총회 및 학술발표회
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    • pp.45-46
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    • 1991
  • 분리막 공정은 상변화를 일으키지 않기 때문에 장치비가 저렴하며, 열 및 화학적으로 민감한 물질, 특히 생체물질 등을 다루는데 적합하다. 또한 최종 생성물질을 정제하는 방법으로는 이온교환 크로마토그래피, 전기 영동 및 친화성 크로마토그래피(affinity chromatography)가 있다. 이중 친화성 크로마토그래피는 가장 선택적이고, 단일 장치로서 기존의 분리방법에 비해 약 1000배 가까이 정제할 수 있다. 본 실험에서는 트립신 억제제인 m-aminobenzamidine을 포함하고 있는 수용성 고분자를 제조하여 트립신을 선택적으로 분리하고자 하였으며, 본 연구 필요한 분리막은 막제조가 용이하며 비용이 저렴한 cellulose acetate막을 사용하였다. CA 막의 세공크기는 겔화매질의 에탄올 농도로 조절하였다. Casting 용액의 조성은 다음 표에 간략하게 나타내었다.

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Determination of Trace Anions in Concentrated Hydrogen Peroxide by Direct Injection Ion Chromatography with Conductivity Detection after Pt-Catalyzed On-Line Decomposition

  • 김도희;이보경;이동수
    • Bulletin of the Korean Chemical Society
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    • 제20권6호
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    • pp.696-700
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    • 1999
  • A method has been developed for the determination of trace anion impurities in concentrated hydrogen peroxide. The method involves on-line decomposition of hydrogen peroxide, ion chromatographic separation and subsequent suppressed-type conductivity detection. H2O2 is decomposed in Pt-catalyst filled Gore-Tex membrane tubing and the resulting aqueous solution containing analytes is introduced to the injection valve of an ion chromatograph for periodic determinations. The oxygen gas evolving within the membrane tubing escapes freely through the membrane wall causing no problem in ion chromatographic analysis. Decomposition efficiency is above 99.99% at a flow rate of 0.4mL/min for a 30% hydrogen peroxide concentration. Analytes are quantitatively retained. The analysis results for several brands of commercial hydrogen peroxides are reported.

홍색 유황세근 Thiocapsa roseopersicina로 부터 생산되는 Hydrogenase의 각종 크로마토그래피에 의한 정제 (Purification of Hydrogenases from Purple Sulfur Bacterium Thiocapsa roseopersicina Using Various Applications of Chromatography)

  • 최은혜;오유관;김미선
    • 한국수소및신에너지학회논문집
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    • 제19권2호
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    • pp.124-131
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    • 2008
  • Crude cytoplasmic fraction of phototrophic purple sulfur bacterium, Thiocapsa roseopersicina NCIB 8347, were initially prepared and purified by sonication, ultracentrifugation, ammonium sulfate fractionation and heat-treatment and it has been previously reported. Using various applications of chromatography far the purification of membrane-bound and soluble hydrogenases from heat-treated enzyme fraction were studied at present report. When the heat-treated enzyme preparation was applied to the anion column chromatography using Q-sepharose, Fraction I and II, which were extracted with the KCl 0-0.5 M gradient, showed the specific evolution hydrogenase activity 3.86 and 2.27 U/mg-protein respectively. Specific hydrogenase activitys of Fraction I and II were further increased to 4.35 and 7.46 U/mg-protein for Fraction I and to 2.49 and 4.41 U/mg-protein fur Fraction II respectively, when hydrophobic interaction column, Phenyl superose, and anion exchange column, Mono-Q, were applied. Size exclusion chromatography using superdex 200 concentrated the hydrogenase Fraction I and II to 9.19 and 7.84 U/mg-protein respectively at the final step of purification.

Apoptosis-Inducing Activity of HPLC Fraction from Voacanga globosa (Blanco) Merr. on the Human Colon Carcinoma Cell Line, HCT116

  • Acebedo, Alvin Resultay;Amor, Evangeline Cancio;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권2호
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    • pp.617-622
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    • 2014
  • Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MPI fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival.

Membrane Lipids of a Marine Ciliate Protozoan Uronema marinum

  • Seo Jung Soo;Kim Ki Hong;Lee Hyung Ho;Chung Joon Ki
    • Fisheries and Aquatic Sciences
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    • 제6권3호
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    • pp.155-159
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    • 2003
  • Lipid composition and fatty acid composition were characterized in the membrane of a marine ciliate protozoan (Uronema marinum). Phospholipids accounted for 70% of total lipid, and the remainder was neutral lipids. Total phospholipids were separated as phosphatidylcholine $(24.26\%)$, phosphatidylethanolamine $(22.21\%)$, phosphatidylinositol $(6.14\%)$, phosphatidyl­serne $(5.11\%)$, cardiolipin $(3.07\%)$ and unidentified phospholipids $(28.72\%)$ through high performance liquid chromatography (HPLC). Fatty acid composition of neutral lipids and phospholipids was determined by gas chromatography (GC), based solely on comparision of retention times. In neutral lipids, the most abundant fatty acid group was monounsaturated fatty acid $(48.3\% of total fatty acids)$ with oleic acid (18:1) and nervonic acid (24:1). Saturated fatty acids comprised $29.6\%$ of total fatty acids, with palmitic acid (16:0), stearic acid (18:0) ane myristic acid (14:0), and polyunsaturated fatty acid accounted for $33.0\%$ with $Di-homo-\gamma-linolenic$ acid (20:3) and linoleic acid (18:2). Wherease phospholipids predominantly contained the fatty acid group in the following order: polyunsaturated fatty acids $(52.7\%\;of\;total\;fatty\;acids)$ with linoleic acid (18:2) and $\gamma-linolenic$ acid (18:3) > monounsaturated fatty acids $(28.5\%\;of\;total\;fatty\;acids)$ with oleic acid (18:1) and palmitoleic acid (16:1) > saturated fatty acids $(25.5\%\;of\;total\;fatty\;acids)$ with palmitic acid (16:0), stearic acid (18:0) and myristic acid (14:0).

Outer Membrane Vesicles Derived from Salmonella Enteritidis Protect against the Virulent Wild-Type Strain Infection in a Mouse Model

  • Liu, Qiong;Yi, Jie;Liang, Kang;Zhang, Xiangmin;Liu, Qing
    • Journal of Microbiology and Biotechnology
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    • 제27권8호
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    • pp.1519-1528
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    • 2017
  • Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

Characteristics of dissolved gases separated from water mixed with exhalation gases without using a compressor

  • Heo, Pil Woo
    • Journal of Advanced Marine Engineering and Technology
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    • 제40권10호
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    • pp.916-921
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    • 2016
  • It is possible for humans to breathe underwater using dissolved oxygen. However, unlike fish, humans need large amounts of oxygen to breathe underwater. Water generally contains small amounts of dissolved oxygen. To get enough dissolved oxygen from water, great volumes of it should be supplied into a separation device. If exhalation gases are used, the amounts of water supplied into the membrane can be decreased. However, the characteristics of exhalation gases after passage through the separation device need to be investigated. To reuse the exhalation gases, the concentration of carbon dioxide should be decreased. A compressor is needed to supply the exhalation gases because of the high pressure generated in the membrane inlet. However, compressors require a lot of power and are heavy, so it is not proper to get the portable separation device. A system without the compressor is needed. If the pressure of the position mixed from the exhalation is less than atmosphere, the compressor is not needed. In this thesis, characteristics of the gases which are mixed with exhalation gases and separated from water after passing the membrane are investigated. The compositions of carbon dioxide, oxygen, and nitrogen are measured with the gas chromatography. The effects of water and exhalation gas flow rates on characteristics of gases separated from water after the membrane are showed.