• Title/Summary/Keyword: gel fraction

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Partial Purification of Protein Kinase C in Glycine max (대두 유식물에서 Protein Kinase C의 부분 정제)

  • 최윤희
    • Journal of Plant Biology
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    • v.36 no.2
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    • pp.171-176
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    • 1993
  • Protein kinase C, a protein related in PI cascade, was partially purified from the cytosol protein of etiolated plants of Glycine max by DEAE-52 cellulose chromatography and phenylsepharose chromatography. When the DEAE column was eluted with 0-0.8 M linear gradient KCl, tow fractions were found that increased the phosphorylation of histon H1 about five and nine-fold in the presence of 5 $\mu\textrm{g}$/mL phosphatidylserine and 0.5 $\mu\textrm{g}$/mL diolein, respectively. These fractions were used as DEAE pool. The reaction eluted with relatively high concentration of KCl was loaded on phyenylsepharose column with 5 mM CaCl2 and eluted with 1 mM EGTA. A fraction contained the protein kinase C, which increased the phosphorylation of the histon H1 was fractionated. To determine the molecular weight of PKC, the fraction eluted from phenylsepharose column was analyzed by 5~15% polyacrylamide gel electrophoresis after concentrated with the Amicon membrane (YM10). That revealed two bands corresponding to 60 and 65 kGy by silver staining of the gel, respectively.

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Purification and Characterization of Angiotensin I-Converting Enzyme Inhibitors from Sinapis alba L.

  • Yuk, Jin-Su;Lim, Young-Hee;Cho, Hong-Yon
    • Preventive Nutrition and Food Science
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    • v.5 no.2
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    • pp.75-80
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    • 2000
  • To separate ACE inhibitors from edible plants, spices, and herbs, 285 extracts of 95 sources were screened for ACE inhibitory activity. The extract of Sinapis alba L. had the most potent ACE inhibitory activity. Mustard seeds were crushed homogeneously and extracted with hexane and water successively. Lyophilized water extract was fractionated with $H_2O$:butanol(1:1). The ACE inhibitor was purified from butanol fraction by methanol precipi-tation, gel filtration, HPLC, and FPLC with Superdex peptide HQ 10/30 column. The active fraction has been purified to homogeneity, which was proven by gel filtration using FPLC system. The yield was 0.02%. The com-pound has a molecular weight of about 640. The compound competitively inhibited ACE activity and the $IC_{50}$ value was 79$\mu\textrm{g}$/ml. The purified compound showed uterus contraction activity in isolated rat uterus.

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Antimutagenic Compounds Indentified from Chloroform Fraction of Persimmon Leaves

  • Moon, Suk-Hee;Jeong-Ok kim;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.203-207
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    • 1996
  • Methanol extract of dried persimmon leaves was refractionated using hexane, chloroform, ethylacetate, n-butanol aqueous fractions. Among these chloroform fraction showed the highest inhibition rate on the mutagenicities of aflatoxin B₁(AFB₁) and 3,2' -dimethyl-4-amino-bipheny1(DMAB) in Salmonella typhimurium TA 100. Chloroform fraction was further fractionated into eight fractions by silica gel column chromatography and thin layer chromatography(TLC). The fraction 5 on TLC exhibited the highest antimutagenic activities in AFB₁and DMAB. 2,4-Decadienal, dihydro-4-methyl-2(3H)-furanone, hexanoic acid 1,4-bis(1-methy1 ethy1)benzene, heptanoic acid, phenol, octanoic acid, nonanoic acid and benzoic acid were tentatively identified from this antimutagenic fraction by GC-MS.

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Antibacterial Activity and Identification of the Active Compound from Tumeric Extract (울금색소 추출물의 항균성과 물질분리 동정)

  • 한신영;최석철
    • Textile Coloration and Finishing
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    • v.14 no.1
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    • pp.11-17
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    • 2002
  • To identify the antimicrobial activity, of tumeric and its active compound tumeric was fractionated into four groups dichloromethane extract, hexane fraction, methanol soluble fraction and residue's extract. They were tested for antibacterial activity against E. coil and S. aureus and the methanol soluble fraction was found lo be the most active fraction. Compound I, II and III were isolated from TLC and silica gel column chromatography in the methanol soluble fraction. These compounds were analyzed by $^1H-NMR\;and\;^{13}C-NMR$ spectra and identified as curcumin I, II and III. They were also tested for antimicrobial activity against E. coli and S. aureus. Curcumin I was the must active curcuminoids due to the phenolic and methoxyl$(OCH_3)$ moiety in the same molecular structure.

삼백초 지상부의 ROS(Reactive oxygen species) 소거효과 및 물질분리

  • 김윤미;김영찬;정신교
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.110.2-111
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    • 2003
  • 삼백초 지상부를 탈지후 60% acetone으로 추출하고 이를 극성에 따라 diethyl ether, ethyl acetate, aqueous fraction과 같이 용매분획하였다. 각 fraction의 superoxide anion, hydrogen peroxide, hydroxyl radical, DPPH radical 에 대한 소거활성을 측정하였다. Diethyl ether fraction의 ROS 소거효과가 비교적 강하였으며 superoxide anion 소거활성은 100ppm에서 86.88%였고, hydroxyl radical 소거활성은 1ppm에서 85.3%로 나타났다. Hydrogen peroxide, DPPH radical 소거활성은 ethyl acetate fraction이 강하게 나타났다. Diethyl ether fraction으로부터 silica gel column chromatography, sephadex LH-20 column chromatography, preparative HPLC에 의하여 수 종의 성분을 분리하였으며 현재 구조동정과 이들의 LDL산화억제 효과 시험 중에 있다.

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Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract (Eisenia bicyclis 에탄올 추출물로부터 분리한 Dieckol의 Lipase 저해 Mode)

  • Jung, Seul-A;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;Cho, Ji-Young;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.112-118
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    • 2013
  • This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

Production of Angiotensin I Converting Enzyme Inhibitory Peptides from Bovine Blood Plasma Proteins (도축 폐혈액 단백질로부터의 Angiotensin I Converting Enzyme 저해 펩타이드의 생산)

  • Hyeon, Chang-Gi;Sin, Hyeon-Gil
    • KSBB Journal
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    • v.14 no.5
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    • pp.600-605
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    • 1999
  • For the production of angiotensin I converting enzyme inhibitory peptides as a material for antihypertensive functional foods from animal blood produced in slaughterhouse, the optimum condition for enzymatic hydrolysis to yield a peptide fraction of the highest activity were investigated with a respect of industrial production. Among several industrially-usable enzymes tested, $Alcalase^?$ produced hydrolysates of the highest activity from total plasma and purified albumin. $IC_50$ values of albumin hydrolysate and its third fraction separated by gel chromatography were 0.5 and 0.02 mg/mL, respectively. The fraction was found to be obtained by a simple ultrafiltration using a membrane of MW cutoff 1,000. The possibility for the industrial production of antihypertensive peptides from animal blood plasma protein was suggested.

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Antimicrobial Activity of Areca catachu L. Extract of against Intestinal Pathogens (장내 유해세균에 대한 빈랑 추출물의 항균 활성)

  • 이갑상;김성효;전승호;박성수;박정순;신용서
    • The Korean Journal of Food And Nutrition
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    • v.11 no.1
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    • pp.36-40
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    • 1998
  • Areca catachu L. which was showed antimicrobial activity against intestinal pathogens through screening herbs related treatments of intestinal diseases, were extracted by methanol and fractionated by n-hexane, ethylether, ethylacetate, and water. Antimicrobial activities of the methanol extract and each fractionates were then investigated under the anaerobic broth system. All fractions of Areca catachu L. except of n-hexane showed antimicrobial activity against all tested pathogens. Especially, ethylacetate fraction of them had the most significant inhibition activity. There is no significant difference of antimicrobial activity among each fractionates. Fraction of Areca catachu L. ethylacetate fractionate, which were fractionated by Sephadex G-200 and Silica gel column chromatography revealed the strongest antimicrobial activity at 5 to 7 and 20 of fraction number, respectively.

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Cytotoxic Constituents from the Forsythiae Fructus against L1210 and HL60 cells (L1210 및 HL60 Cell에 대한 연교의 세포독성 성분)

  • Lee, Jun-Seong;Min, Byeong-Seon;Bae, Gi-Hwan
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.462-467
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    • 1996
  • Forsythiae Fructus was studied on cytotoxic activities for the purpose of finding out active consituents against L1210 and HL60 cells. To isolate the active ones, the methanolic extract was partitioned into water insoluble and water soluble fractions. Furthermore, the water soluble fraction was fractionated into four parts, n-hexane, benzene, ethylacetate and water fractions. Among these, the water insoluble fraction showed the most potent cytotoxic activities on L1210 and HL60 cells in vitro. The water insoluble fraction was applied to silica gel column chromatography and divided into 5 fractions(fr. 1-5). The active constituents I and II were isolated from fr.2 and 3, respectively, by repeated silica gel column chromatography and recrystallization. The constituents were identified as 3${\beta}$-acetylbetulinic acid and betulinic acid by means of physicochemical data. The $ED_{50}$ values of 3${\beta}$-acetylbetulinic acid and betulinic acid were 9.10 and 16.43${\mu}g$/ml against L1210 cells and 2.72 and 2.41${\mu}g$/ml against HL60 cells, respectively.

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Hematopoietic Characteristics of the Ethylacetate Fraction of Panax ginseng and Identification of the Chemical Constituents (인삼의 에칠아세테이트 추출분획의 조혈활성 특성 및 구성 성분 동정)

  • Park, Jong-Dae;Wee, Jae-Joon;Kim, Man-Wook;Lee, Hyong-Joo
    • Applied Biological Chemistry
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    • v.32 no.2
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    • pp.137-142
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    • 1989
  • The ethylacetate soluble fraction of Panax ginseng extract, which has been reported to exhibit hematopoietic effect, was divided into three subfractions, i.e., Fr. I, Fr. II and Fr. III by silica gel column chromatography. The hematopoietic effect of each subfraction was examined in rats treated with anticancer agent, Ara-C($1-{\beta}-D-arabinofuranosylcytosine)$). Among them, Fr. III showed 90% recovery of leucocyte and erythrocyte counts in bonemarrow cell depleted rats, suggesting it to be hematopoietic fraction. From Fr. III, two major compounds were isolated and identified as ginsenoside $Rh_1$ and $Rg_2$ by $^{13}C-NMR$ and degradation method.

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