• Title/Summary/Keyword: Isolation Technology

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Identification and quantification of major malonyl ginsenosides isolated from Panax ginseng C.A. Meyer (인삼(Panax ginseng C.A. Meyer)로부터 Malonyl ginsenoside의 분리 및 정량분석)

  • Shin, Woo Cheol;Jung, Jiyun;Na, Hyeon Seon;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Choi, Bo-Ram;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Yi;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.375-384
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    • 2019
  • The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H2O fractions using Diaion HP-20. The repeated SiO2 or octadecyl SiO2 column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Glycerides from the Aerial Parts of Garland (Chrysanthemum coronarium L.) and Their Inhibitory Effects on ACAT, DGAT, FPTase, and $\beta$-Secretase

  • Song, Myoung-Chong;Yang, Hye-Joung;Cho, Jin-Gyeong;Chung, In-Sik;Kwon, Byoung-Mog;Kim, Dae-Keun;Baek, Nam-In
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.95-102
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    • 2009
  • The aerial parts of garland (Chrysanthemum coronarium L.) were extracted in 80% aqueous methanol (MeOH) and the concentrated extract was then partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. EtOAc and n-BuOH fractions resulted in 4 glycerides with the application of octadecyl silica gel and silica gel column chromatography. The chemical structures of the glycerides were determined using several spectroscopic methods, including nuclear magnetic resonance (NMR) and mass spectrometry (MS) as (2S)-1-O-palmitoyl-sn-glycerol (1), (2S)-1-O-oleoyl-2-O-oleoyl- 3-O-$\beta$-D-galactopyranosyl-sn-glycerol (2), (2S)-1-O-palmitoyl-2-O-linoleoyl-3-O-phosphorouscholine-sn-glycerol (3), and (2S)-1-O-linolenoyl-2-O-palmitoyl-3-O-[$\alpha$-D-galactopyrasyl-($1{\rightarrow}6$)-$\beta$-D-galactopyranosyl]-sn-glycerol (4). The free fatty acids of these glycerides were determined with gas chromatography (GC)-MS analysis following alkaline hydrolysis and methylation. These glycerides demonstrated an inhibitory effect on acyl-CoA: cholesterol acyltransferase (ACAT, compound 1: $45.6{\pm}0.2%$ at $100{\mu}g/mL$), diacylglycerol acyltransferase (DGAT, compound 1: $59.1{\pm}0.1%$ at $25{\mu}g/mL$), farnesyl protein transferase (FPTase, compound 2: $98.0{\pm}0.1%$; compound 3: $55.2{\pm}0.1%$ at $100{\mu}g/mL$), and $\beta$-secretase ($IC_{50}$, compound 4: $2.6{\mu}g/mL$) activity. This paper is the first report on the isolation of these glycerides from garland and their inhibitory activity on ACAT, DGAT, FPTase, and $\beta$-secretase.

Cloning and Expression of a Human Homolog of Mouse Gamml, MVGI, Localized in 12q13 (인간염색체 12q13에 내재한 마우스 Gamm1의 인간유전자 homolog, MYG1의 클로닝과 발현)

  • Yang, Keum-Jin;Lee, Hyoung-Nam;Bae, Youn-Jung;Shin, Dong-Jik;Kim, Eun-Min;Yoon, Jong-Bok;Park, Young-Il;Kim, Jun;Yu, Ji-Chang;Kim, Sung-Joo
    • KSBB Journal
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    • v.17 no.4
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    • pp.370-375
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    • 2002
  • Isolation of a gene and determination of its expression pattern are essential in understanding its function. Among the genes localized in 12ql3, stSG3435 EST was chosen to study its expression pattern. The full-length CDNA was cloned by screening of human brain CDNA library and its sequence was determined by serial deletion followed by automated sequencing of the clones with overlapping fragments. The sequence analysis revealed that stSG 3435 CDNA displayed 100% identity to human MYGI and 86% identity to mouse melanocyte proliferation gene-1 (Gamm 1) originally identified from melanocyte, suggesting that MYGI determined by Northern blot analysis revealed the strongest expression in testes with ubiquitous expression in all the tissues tested. In order to investigate the cellular localization of its protein product, the green fluorescence protein gene was fused into the full-length coding sequence of MYGI, Transfection of the fusion construct followed by confocal microscopy resulted in the green fluorescence signal as a punctate state in cytoplasm indication that MYGI was localized in one of the cellular organelles.

MANAGING A PROLONGED STATION BLACKOUT CONDITION IN AHWR BY PASSIVE MEANS

  • Kumar, Mukesh;Nayak, A.K.;Jain, V;Vijayan, P.K.;Vaze, K.K.
    • Nuclear Engineering and Technology
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    • v.45 no.5
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    • pp.605-612
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    • 2013
  • Removal of decay heat from an operating reactor during a prolonged station blackout condition is a big concern for reactor designers, especially after the recent Fukushima accident. In the case of a prolonged station blackout condition, heat removal is possible only by passive means since no pumps or active systems are available. Keeping this in mind, the AHWR has been designed with many passive safety features. One of them is a passive means of removing decay heat with the help of Isolation Condensers (ICs) which are submerged in a big water pool called the Gravity Driven Water Pool (GDWP). The ICs have many tubes in which the steam, generated by the reactor core due to the decay heat, flows and condenses by rejecting the heat into the water pool. After condensation, the condensate falls back into the steam drum of the reactor. The GDWP tank holds a large amount of water, about 8000 $m^3$, which is located at a higher elevation than the steam drum of the reactor in order to promote natural circulation. Due to the recent Fukushima type accidents, it has been a concern to understand and evaluate the capability of the ICs to remove decay heat for a prolonged period without escalating fuel sheath temperature. In view of this, an analysis has been performed for decay heat removal characteristics over several days of an AHWR by ICs. The computer code RELAP5/MOD3.2 was used for this purpose. Results indicate that the ICs can remove the decay heat for more than 10 days without causing any bulk boiling in the GDWP. After that, decay heat can be removed for more than 40 days by boiling off the pool inventory. The pressure inside the containment does not exceed the design pressure even after 10 days by condensation of steam generated from the GDWP on the walls of containment and on the Passive Containment Cooling System (PCCS) tubes. If venting is carried out after this period, the decay heat can be removed for more than 50 days without exceeding the design limits.

Isolation of Microorganisms and Development of Microbial Augmentation for Treatment of Paper Mill Wastewater (제지폐수 처리용 미생물의 분리 및 복합 미생물제제의 개발)

  • Kang, Dae-Ook;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.554-560
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    • 2011
  • This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.

Effect of Phenobarbital on the Hepatic Clearance of Diltiazem in Isolated Rat Hepatocytes (흰쥐 분리 간세포에 있어서 딜티아젬의 간클리어런스에 미치는 페노바르비탈의 영향)

  • Lee, Yong-Bok;Oh, Joon-Kyo;Kho, Ik-Bae
    • Journal of Pharmaceutical Investigation
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    • v.26 no.1
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    • pp.33-41
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    • 1996
  • In order to study the effect of phenobarbital(PB) on the hepatic transport of diltiazem(DTZ), $Ca^{2+}$ channel blocker, we used isolated hepatocytes of rat which was intraperitoneally pretreated with phenobarbital sodium(75 mg/kg) for four days once a day. For the isolation of rat liver cells, a modification of the two step procedure of Seglen was used. DTZ was dissolved in incubation buffer to the final DTZ concentrations of 200, 400, 600, 800 and 1000 ng/ml in order to elucidate the uptake characteristics of DTZ by hepatocytes. Reactions were stopped at 10, 20, 30, 45, 60, 90, 120 and 300 sec. The initial velocity was determined by disappearance of diltiazem in the hepatocyte suspension. On the other hand, to determine the effect of PB on the in vitro hepatic intrinsic clearance of DTZ we obtained the metabolism rates of DTZ in the control and the PB-pretreated rat hepatocyte at various time intervals. According to pretreatment with PB, the size of hepatocyte and the amount of protein per $10^6$ cells were significantly (p<0.01) increased from $26.92{\pm}0.1364\;m$ to $35.31{\pm}1.00\;m$ and from $468{\pm}6.5\;{\mu}g/10^6$ cells to $628.8{\pm}12.1{\mu}g/10^6$ cells, respectively. In the case or hepatic uptake of diltiazem, $K_m$ was not different in the normalization by cell numbers and increased from $2.90\;{\mu}M\;to\;13.89\;{\mu}M$ in the normalization by protein amount. $V_max$ was increased regardless of normalization by protein amount and cell numbers, from $1.21\;{\mu}mole/min \;{\cdot}\;mg\;protein\;to\;3.96\;{\mu}mole/min\;{\cdot}\;mg\;protein\;and\;from\;2.38\;{\mu}mole/min\;{\cdot}\;10^6\;cells\;to\;2.83\;{\mu}mole/min\;{\cdot}\;10^6\;cells$, respectively. The in vitro hepatic intrinsic clearance of DTZ was significantly (p<0.01) increased from $0.640{\pm}0.038\;ml/mim\;{\cdot}\;10^6\;cells\;to\;2.385{\pm}0.212\;ml/min\;{\cdot}\;10^6\;cells$ due to PB-pretreatment. These results suggest that the uptake of DTZ by hepatocyte is extremely fast and PB enhances the hepatic intrinsic metabolic clearance of DTZ.

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Constituents of Antimutagenic Factor from Brown Rice (현미의 항돌연변이 활성물질의 구성성분)

  • Chun, Hyang-Sook;Moon, Tae-Wha;Kim, In-Ho
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.478-483
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    • 1995
  • To investigate the constituents of antimutagenic factor from brown rice, methanol extracts were fractionated into ether, ethylacetate, buthanol and aqueous fractions. The ether fractions showed distinct antimutagenic effect and active spot were selected by silica gel chromatography. The specific activity of active spot decreased with isolation of the active components from the methanol extract. Qualitative analyses of the active spot by using various spraying reagents revealed that ninhydrin and orcinol did not develop colored reactions. But, ferric chloride, 2,7-dichlorofluorescein, antimony pentachloride, phosphomolybdic acid, bromothymol blue and rhodamine 6G led to colored reactions. These results suggested that the consitituents of active material were neither polar nor nitrogen-containing compounds and that they may contain phenolic compounds and fatty acid derivatives. Main compounds of the active spot were analyzed to be o-hydroxy benzyl alcohol(saligenin), octanoic acid(caprylic acid), 9,12-cis-octadecadienoic acid(linoleic acid), 11-cis-octadecenoic acid(oleic acid), hexadecanoic acid(palmitic acid), 1H-indole-2-carboxylic acid and 1,2-benzenedicarboxylic acid(phthalate) in GC/Mass spectrum, and antimutagenicity of these active compounds using standard regeant was reconfirmed in S. typhimurium reversion assay.

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Inhibitory Substance Produced by Aspergillus sp. on the Snake Venom Proteinase - Isolation of Microorganism and Biological Activities of the Inhibitor - (Aspergillus 속 균주가 생성되는 사독 Proteinase에 대한 저해물질 - 균의 분리 및 저해물질의 생물학적 작용상 -)

  • Hyun, Nam-Joo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.129-134
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    • 1987
  • Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, $\alpha$-chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but not stable at $65^{\circ}C$ in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10$0^{\circ}C$ for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

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Physiological and Ecological Characteristics of Hemolytic Vibrios and Development of Sanitary Countermeasure of Raw Fisheries Foods 1. Isolation and Identification of Novel Pathogenic Vibrio sp. Producing Hemolysin (용혈독소를 생산하는 기수성 비브리오균의 생리${\cdot}$생태적 특성과 수산식품의 위생 대책 1. 용혈독소를 생산하는 새로운 병원성 Vibrio sp.의 분리와 동정)

  • KIM Young-Man;CHOI Gil-Bae;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.361-366
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    • 1997
  • To determine the physiological, biochemical characteristics and toxicity of hemolysin produced by a novel sucrose positive Vibrio (Vibrio sp. D5) isolated from estuary of Kum river, it was compared with already known sucrose positive Vibrio. Salinity, pH, temperature and conductivity of place where Vibrio sp. D5 was isolated were $4.7\%_{\circ},\;7.6,\;24^{\circ}C$ and $7800{\mu}MHOS$, respectively. Physiological and biochemical characteristics distingiushed Vibrio sp. D5 from other sucrose positive Vibrio: V. alginoipicus, V. cholerae, V. cincinnatiensis, V. fluvialis, V. furnissii and V. metschnikovii. The range of salinity and pH for growth of Vibrio sp. D5 were $0.5\%\~7.5\%$ and $4.5\~9.5$, respectively. Vibrio sp. D5 exhibited typical yellow colony on TCBS agar plate and curved rod type upon transmission electron microscopy (TEM). Vibrio sp. D5 had lethal toxicity against mouse in case of intraperitoneal injection with its culture and showed hemolysin activity on human blood agar and sheep blood agar. Ubrio sp. D5 also demonstrated vascular permeability activity toward rat. From the above results, Vibrio sp. D5 was ascertained to be a novel pathogenic Vibrio.

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Fabrication, Estimation and Trypsin Digestion Experiment of the Thermally Isolated Micro Teactor for Bio-chemical Reaction

  • Sim, Tae-Seok;Kim, Dae-Weon;Kim, Eun-Mi;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Hyup;Kim, Yong-Kweon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.5 no.3
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    • pp.149-158
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    • 2005
  • This paper describes design, fabrication, and application of the silicon based temperature controllable micro reactor. In order to achieve fast temperature variation and low energy consumption, reaction chamber of the micro reactor was thermally isolated by etching the highly conductive silicon around the reaction chamber. Compared with the model not having thermally isolated structure, the thermally isolated micro reactor showed enhanced thermal performances such as fast temperature variation and low energy consumption. The performance enhancements of the micro reactor due to etched holes were verified by thermal experiment and numerical analysis. Regarding to 42 percents reduction of the thermal mass achieved by the etched holes, approximately 4 times faster thermal variation and 5 times smaller energy consumption were acquired. The total size of the fabricated micro reactor was $37{\times}30{\times}1mm^{3}$. Microchannel and reaction chamber were formed on the silicon substrate. The openings of channel and chamber were covered by the glass substrate. The Pt electrodes for heater and sensor are fabricated on the backside of silicon substrate below the reaction chamber. The dimension of channel cross section was $200{\times}100{\mu}m^{2}$. The volume of reaction chamber was $4{\mu}l$. The temperature of the micro reactor was controlled and measured simultaneously with NI DAQ PCI-MIO-16E-l board and LabVIEW program. Finally, the fabricated micro reactor and the temperature control system were applied to the thermal denaturation and the trypsin digestion of protein. BSA(bovine serum albumin) was chosen for the test sample. It was successfully shown that BSA was successfully denatured at $75^{\circ}C$ for 1 min and digested by trypsin at $37^{\circ}C$ for 10 min.