• Title/Summary/Keyword: Solvent fractionation

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Antithrombin and Thrombosis Prevention Activity of Buckwheat Seed, Fagopyrum esculentum Moench (메밀 종자의 항트롬빈 활성과 혈전증 예방효과)

  • Sohn, Ho-Yong;Kwon, Chong-Suk;Son, Kun-Ho;Kwon, Gi-Seok;Ryu, Hee-Young;Kum, Eun-Joo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.2
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    • pp.132-138
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    • 2006
  • Direct thrombin inhibitor, which is effective to prevent or cure the thrombosis, has been investigated in worldwide. In this study, we tried to screen antithrombosis agent from edible or medicinal plant. A strong antithrombin activity was identified from methanol or $95\%$ ethanol extract of buckwheat seeds. The solvent fractionation of buckwheat extracts using hexane, ethylacetate, butanol revealed that the butanol fraction has a prominent antithrombin activity. Thrombin time (blood-clot formation time) exceeded to over $2,000\%$ by addition of the butanol fraction at concentration of $312.5{\mu}g/mL$, whereas thrombin time extended to $336\%$ by addition of aspirin at concentration of $1,500{\mu}g/mL$. The butanol fraction showed anthrone-positive and ninhydrine-negative reaction. The active components were heat-liable, acid-unstable non-proteinous macromolecules (>30 KD). In vivo analysis using ICR male mouse showed that the buckwheat extract was superior than the aspirin in pulmonary thrombosis, KCN-induced coma and death. Our results suggest that the buckwheat is a potential as an antithrombosis agent and medicinal food.

Search for Plant-originated Antibacterial Compounds Against Pathogen (Acidovorax avenae subsp. citrulli) of Watermelon Bacterial Fruit Blotch (수박 과실썩음병 병원균(Acidovorax avenae subsp. citrulli)에 대한 식물유래 항균 활성물질 탐색)

  • Noh, Jin-Taek;Choi, Yong-Hwa
    • Korean Journal of Organic Agriculture
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    • v.23 no.1
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    • pp.77-89
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    • 2015
  • 133 Species of medicinal plants were used for the development of natural agrichemicals with anti-microbial activity against Acidovorax avenae subsp. citrulli, a pathogen of bacterial fruit blotch in watermelon. The MeOH-extracts of these medicinal plants were examined for anti-microbial activity by bioassay. The MeOH-extract of Citrus unshiu Markovich had the strongest antibacterial activity against Acidovorax avenae subsp. citrulli. To identify anti-microbial compounds from Citrus unshiu Markovich, solvent-fractionation was used. The fraction of hexane, which showing the highest value of anti-microbial activity, was analyzed by GC-MS. Each mass spectra, corresponding to each peak of chromatogram, was compared to mass database of Wiley library. As a result, d-Limonene, ${\gamma}$-terpinene, ${\beta}$-linalool, terpineol, palmitic acid, 9,12-octadecadienoic acid, Linolenic acid, and stigmasterol were identified. Among them, d-Limonene, ${\gamma}$-terpinene, ${\beta}$-linalool, and terpineol confirmed to be shown the anti-microbial activity by bioassay. Especially, d-Limonene and ${\gamma}$-terpinene found to have strong activity. In conclusion, we thought d-limonene and ${\gamma}$-terpinene from Citrus unshiu Markovich. Latin, had anti-microbial activity against Acidovorax avenae subsp. citrulli and could be candidates for the control agents for the control of bacterial fruit blotch in watermelon.

Thrombin Inhibition Activity of Fructus Extract of Crataggus pinnatifida Bunge (산사자 추출물의 트롬빈 저해활성)

  • Ryu, Hee-Young;Kim, Yung-Kwan;Kwun, In-Sook;Kwon, Chong-Suk;Jin, Ing-Nyol;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.535-539
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    • 2007
  • The fructus of Crataegus pinnatifida Bunge has been used as medicinal and food source in worldwide. In this study, a strong direct thrombin inhibition and antithrombosis activity were identified from the methanol extract of C. pinnatifida Bunge fructus. The solvent fractionation of fructus extract using hexane, ethylacetate, butanol revealed that the butanol fraction has a prominent antithrombin activity. Thrombin time(blood-clot formation time) and activated partial thromboplastin time(aPTT) extended to 835% and 315% by addition of the butanol fraction at concentration of 1.25 mg/mL, whereas thrombin time extended to 287% by addition of aspirin at concentration of 1,25 mg/mL. The butanol fraction showed anthrone-positive and weak ninhydrine-postive reaction. The thrombin inhibitory activity was not related to previously reported flavonoids or polyphenols. The activity was maintained against acid treatment(0.5 N HCl for 120 min), but rapidly lost by heat-treatment($100^{\circ}C$ for 30 min). Our results suggested that fructus of C. pinnatifida Bunge with non-heat treatment process could be developed as a natural source of antithrombosis.

Biosynthesis of Copolyesters Consisting of 3-Hydroxyvalerate and Medium-chain-length 3-hydroxyalkanoates by the Pseudomonas aeruginosa P-5 Strain (Pseudomonas aeruginosa P-5 균주로부터 3-Hydroxyvalerate와 Medium-chain-length 3-hydroxyalkanoates로 구성된 공중합체의 생합성)

  • Woo, Sang-Hee;Kim, Jae-Hee;Ni, Yu-Yang;Rhee, Young-Ha
    • Korean Journal of Microbiology
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    • v.48 no.3
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    • pp.200-206
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    • 2012
  • A bacterial strain capable of synthesizing polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units was isolated from activated sludge using the enrichment culture technique. The organism, identified as Pseudomonas aeruginosa P-5, produced polyesters consisting of 3-hydroxyvalerate and medium-chain-length (MCL) 3-hydroxyalkanoate monomer units when $C_{-odd}$ alkanoic acids such as nonanoic acid and heptanoic acid were fed as the sole carbon source. Solvent fractionation experiments using chloroform and hexane revealed that the 3-hydroxyalkanoate monomer units in these polyesters were copolymerized. The molar concentration of 3-hydroxyvalerate in the polyesters produced were significantly elevated up to 26 mol% by adding 1.0 g/L valeric acid as the cosubstrate. These copolyesters were sticky with low degrees of crystallinity. The PHA synthase genes were cloned, and the deduced amino acid sequences were determined. P. aeruginosa P-5 possessed genes encoding MCL-PHA synthases (PhaC1 and PhaC2) but lacked the short-chain-length PHA synthase gene, suggesting that the MCL-PHA synthases from P. aeruginosa P-5 are uniquely active for polymerizing (R)-3-hydroxyvaleryl-CoA as well as MCL (R)-3-hydroxyacyl-CoAs.

Anti-oxidant and Anti-inflammation Activity of Fractions from Aster glehni Fr. Schm. (섬쑥부쟁이(Aster glehni Fr. Schm.) 분획물의 항산화 및 항염증 활성)

  • Kim, Han-Hyuk;Park, Gun-Hye;Park, Kang-Soo;Lee, Jin-Young;An, Bong-Jeun
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.434-441
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    • 2010
  • The Plants and their extracts containing polyphenol have been shown to be associated with decreased the cause of aging and variety of disease such as reaction oxygen species (ROS) and reactive nitrogen species (RNS) in several recent studies. We conducted to investigate whether the extracts and fractionation isolated from Aster glehni Fr. Schm. has an inhibitory effect association with oxidation or inflammation. The Aster glehni Fr. Schm. 70% aq. MeOH was fractioned according to polarity with n-hexane layer, EtOAc layer, n-BuOH layer and water layer. The electron donating ability of EtOAc, n-BuOH solvent fraction from Aster glehni Fr. Schm. was about 58.0%, 46.4% at $50\;{\mu}g/mL$, respectively. The superoxide anion radical inhibitory effect of EtOAc extracts was about 64.65% at $50\;{\mu}g/mL$, and n-BuOH extracts was 35.66% at $50\;{\mu}g/mL$. EtOAc layer to the inhibition activity of hyaluronidase and lipoxygenase were inhibited about 24.37%, 29.5% at $5\;{\mu}g/mL$. In the anti-inflammation effect of EtOAc layer inhibited the generation of nitric oxide. also, these results showed that EtOAc extract inhibited 81.5% at $50\;{\mu}g/mL$ on the expressions of iNOS protein in Raw264.7 cell line.

Chemical Composition and Antimicrobial Activities of Houttuynia cordata Thunb. (어성초의 화학성분 및 항미생물 활성)

  • Kim, Keun-Young;Chung, Dong-Ok;Chung, Hee-Jong
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.400-406
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    • 1997
  • Chemical composition and antimicrobial activity of Houttuynia cordata Thunb. were investigated to develop a natural food preservative from it. Aspartic acid, glutamic acid, glycine and arginine were major amino acids of Houttuynia cordata Thunb., but were present in a trace amount. Free sugars were composed of glucose, fructose, sucrose and maltose and major fatty acids were linolenic acid, linoleic acid, oleic acid, and palmitic acid. Mineral elements were potassium, calcium, magnesium, iron, zinc, and copper. Antimicrobial activities were shown in acidic, neutral and phenolic fraction of Houttuynia cordata Thunb., but not in basic fraction. Among the four fractions, neutral fraction showed the strongest antimicrobial activities against microorganisms tested, such as Pseudomnas aeruginosa, Micrococcus luteus, Bacillus subtilis, Aspergillus parasiticus and Aspergillus flavus. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) of the neutral fraction varied according to microoganisms tested. The lowest values of MIC (0.0075 g eq./mL) and MLC (0.10 g eq./mL) were obtained from Pseudomonas aeruginosa.

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Polyacetylene Compounds from Panax ginseng C.A. Meyer (인삼의 Polyacetylene 화합물)

  • Shim Sang Chul;Chang Suk-Ku
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.122-128
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    • 1988
  • Several major polyacetylene compounds were isolated from the petroleum-ether fraction of fresh Korean ginseng roots through solvent fractionation. partition and silica gel column chromatography. Further separation of acetylenic compounds was accomplished by bonded normal phase HPLC utilizing a moderately nonpolar microparticulate column. The preparative separation for the various spectral measurements was carried out by low pressure preparative liquid chromatography. The chemical structure of these polyacetylenes separated was determined by UV. IR/FTIR. $^{1}H$ NMR. mass spectral and elemental analysis. These are identified to be heptadeca-1-en-4.6-diyn-3.9.l0.-triol [1] heptadeca-1.9-dien-4.6-diyn-3-ol. heptadeca-1.8-dien-4.6-diyn-3.10-diol and the 4th was denatured polyacetylene. heptadeca-1.4-dien-6.8-diyn-3.10-diol. Two different p-substituted benzoates of panaxynol were synthesized for the determination of exciton chirality. The circular dichroism spectra in the UV region show that panaxynol p-bromobenzoate and p-dimethyl-aminobenzoate constitute negative exciton chirality [2]. Isolated major polyacetylene compounds were irradiated in aerated solution with 300 nm UV light to obtain the oxidized product at the allylic alcohol center to corresponding carbonyl compounds such as heptadeca-1-en-4.6-diyn-9.10-diol-3-one and heptadeca-1.9-dien-4.6-diyn-3-one. These photooxidation compounds have en-on-diyne chromophore and undergo nucleophilic addition reaction with methanol to yield ${\beta}-methoxy$ carbonyl compounds such as heptadeca-9-en-4.6-diyn-1-methoxy-3-one and heptadeca-4.6-diyn-1-methoxy-9.10-diol-3-one.

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Isolation and Identification of Antioxidants from Makgeolli (막걸리로부터 항산화 화합물의 단리.정제 및 동정)

  • Wang, Seung-Jin;Lee, Hyoung-Jae;Cho, Jeong-Yong;Park, Keun-Hyung;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.14-20
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    • 2012
  • The present study was carried out to isolate and identify of antioxidants from makgeolli. Makgeolli (3 L) was filtered and the residue was extracted by MeOH. The combined filtrates and MeOH extracts were successively solventfractionated by n-hexane, EtOAc, and BuOH. In the antioxidative activity against DPPH and $ABTS^+$ radicals of each fraction obtained after solvent-fractionation, EtOAc and BuOH layers showed higher activities than other fractions. Therefore, the two layers were respectively purified by column chromatography and HPLC. The isolated compounds were subjected to NMR and MS analyses and identified as 4-hydroxybenzaldehyde (1, 2.0 mg), 2-(4-hydroxyphenyl)ethanol (2, tyrosol, 15.3 mg), trans- and cis-ferulic acids (3 and 4, 1.2 mg), 1H-indole-3-ethanol (5, tryptophol, 3.4 mg), dimethyl succinate (6, 14.9 mg), succinic acid (7, 7.4 mg), and mono-methyl succinate (8, 7.8 mg). The presence of 1-5 in makgeolli have never before been reported.

Biosynthesis and Degradation of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in Alcaligenes sp. SH-69 (Alcaligenes sp. SH-69에서의 Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) 생합성 및 분해)

  • Ryu, Kang-Eun;Choi, Gang Guk;Park, Sang Kyu;Kim, YoungBaek;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.219-224
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    • 1998
  • The cyclic metabolism of poly(3-hydroxyhutyrate-co-3-hydroxyvalerate) synthesized from glucose by Alcaligenes sp. SH-69 in the presence or absence of new carbon substrate was investigated. In batch culture, the content and weight average molecular weight of the copolymer already stored in the cell decreased rapidly when there was no other carbon source available. After the depletion of carbon source, the amount of high molecular weight copolymer decreased more rapidly than that of low molecular weight copolymer, and as a result, average molecular weight distribution shifted to the lower value. The addition of a mixture of glucose and levulinic acid when the initial carbon substrate, glucose, was nearly depleted supported the continual increase in cell mass and the accumulation of poly(3HB-co-3HV) with high molar fraction of 3HV. However, solvent fractionation of the polymer with acetone revealed the degradation of pre-existing polyhydroxyalkanoale (PHA) in parallel with the synthesis of PHA from new carbon substrate. Even though PHAs obtained from each substrate alone were the copolymer of 3HB and 3HV, it was found that the polymer accumulated in the cells grown by sequential feeding was mainly physical mixture of two poly(3HB-co-3HV) copolymers containing different molar fractions of 3HV.

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Biochemical Characterization of a Protease with Fibrinolytic Activity from Maggots of Protaetia brevitarsis (Protaetia brevitarsis의 maggot로부터 fibrinolytic activity을 가진 protease의 생화학적 특성 연구)

  • Chang, Jeong-Hyun;Jo, Ji-Young;Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyoung;Kim, Kwang-Youn;Park, Byoung-Keun;Ahn, Soon-Cheol;Kwon, Heun-Young
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.606-612
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    • 2007
  • Fibrin clots remained in blood vessels can be one of the serious factor caused cardiovascular disease, such as ischemia, infarction and necrosis The development of an antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. In this study, the fibirinolytic protease was prepared from the maggots of Protaetia brevitarsis using ammonium sulfate fractionation and desalting column. The optimum pH and temperature for the enzyme activity were pH 9.0 and $50^{\circ}C$, respectively. The enzyme activity was relatively stable at pH 7.0-9.0 and temperature below $60^{\circ}C$. The activity of the enzyme was strongly inhibited by phenylmethanesulfonyl fluoride. And the activity of the enzyme was inhibited by $Ca^{2+}\;and\;Zn^{2+}$, but it was not by $Mg^{2+}\;and\;Fe^{2+}$ ions. In these experimental results, we have speculated that the enzyme derived from maggots of Protaetia hrevitarsis is a serine protease with a strong fibrinolytic activity.