• Title/Summary/Keyword: isolation and purification

Search Result 286, Processing Time 0.035 seconds

Purification and Identification of Antioxidant Compounds from Dolichos lablab L. Seeds (백편두의 항산화 물질 분리 및 동정)

  • Kwon, Nam Woo;Kim, Jae Yeon;Cho, Yong Beom;Hwang, Bang Yeon;Kim, Jun Gu;Woo, Sun Hee;Lee, Moon Soon
    • Korean Journal of Medicinal Crop Science
    • /
    • v.27 no.6
    • /
    • pp.419-426
    • /
    • 2019
  • Background: This study aimed to identify antioxidant compounds from the seeds of Dolichos lablab L. by bioassay-guided isolation and recrystallization. Methods and Results: The water layer of D. lablab L. seed extract inhibits intracellular reactive oxygen species (ROS) expressing the 2',7'-dichlorofluorescein diacetate (DCF-DA), Cu/Zn superoxide dismutase (SOD) and catalase genes, as determined by quantitative real-time PCR (qRT-PCR). Two compounds were purified from the water layer of the seeds of D. lablab L. using column chromatography and prep-high performance liquid chromatography (HPLC). Using nuclear magnetic resonance (NMR) and electrospray Ionization mass spectrometry (ESI-MS), their chemical structures were identified as 5-[(2-acetyl-2,3-dihydro-1H-indazol-1-yl)carbonyl]-4,5-dihydro-3H-furan-2-one (C14H14N2O4) and stachyose. Conclusions: Two active antioxidant compounds were purified from the seed extract of D. lablab L. seed extract and the structures of these compounds were identified as C14H14O4N2 and stachyose.

Low Density Lipoprotein-oxidation Inhibitory Phytochemicals from the Fruits of Rhus parviflora

  • Shrestha, Sabina;Park, Ji-Hae;Cho, Jin-Gyeong;Lee, Dae-Young;Kang, Ji-Hyun;Li, Hua;Jeong, Tae-Sook;Kim Cho, Somi;Lee, Dong-Sun;Baek, Nam-In
    • Journal of Applied Biological Chemistry
    • /
    • v.58 no.2
    • /
    • pp.109-112
    • /
    • 2015
  • Fruits of Rhus parviflora were extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. Purification of EtOAc fraction led to isolation of fifteen polyphenols of which structures were identified by spectroscopic methods including 2D-NMR. Most compounds apart from compound 10 inhibited low density lipoproteinoxidation within $IC_{50}$ value of $10{\mu}M$. Among compounds, taxifolin (2), quercetin 3-O-${\alpha}$-L-rhamnopyranoside (13), agathisflavone (5) sulfuretin (4), and aureusidin (3) showed $IC_{50}$ values 0.9, 0.8, 5.8, 2.9, and $2.4{\mu}M$ which were of highly significant in comparison positive control butylated hydroxytoluene with $IC_{50}$ value of $2.1{\mu}M$. The results indicate fruits of R. parviflora as a source of antihypercholesterolemic compounds.

Antioxidant Activity of the Salt Marsh Plant Corydalis heterocarpa (염주괴불주머니 (Corydalis heterocarpa) 의 항산화 활성)

  • Kim, You-Ah;Lee, Jung-Im;Lee, Jin-Hyeok;Kong, Chang-Suk;Nam, Taek-Jeong;Seo, Young-Wan
    • KSBB Journal
    • /
    • v.24 no.5
    • /
    • pp.469-476
    • /
    • 2009
  • The antioxidant activities of two crude extracts ($CH_2Cl_2$ and MeOH) and their solvent fractions (n-hexane, 85% aq. MeOH, n-BuOH, and $H_2O$ fractions) from Corydalis heterocarpa were determined by evaluating authentic $ONOO^-$ and $ONOO^-$ generated from SIN-1 (3-morpholinsydnonimine) in vitro as well as by measuring the degree of occurrence of intracellular reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of solvent fractions on authentic $ONOO^-$ increased in the order of n-BuOH > 85% aq. MeOH > $H_2O$ > n-hexane fractions, while those on $ONOO^-$ generated from SIN-1 increased in the order of n-BuOH > 85% aq. MeOH > $H_2O$ > n-hexane fractions. In addition, all solvent fractions effectively inhibited the intracellular ROS and NO levels. The n-BuOH fraction especially exhibited the strongest ROS scavenging effect. Further purification of n-BuOH fraction led to the isolation of cnidimoside A, which presented the potent ROS scavenging effect at $10\;{\mu}M$. From these results, extracts of C. heterocarpa and its component, cnidimoside A, were predicted to be potentially useful as ingredients for protecting against oxidation.

Identification of Water Soluble Metabolites of Pentachlorophenol(PCP) in the Suspension Cultures of Soybean and Rice Cells;2. Isolation and characterization of PCP glucose conjugates (콩과 벼 현탁배양시(懸濁培養時) PCP 수용성대사물(水溶性代謝物)의 동정(同定);2. PCP glucose conjugates의 분리(分離) 및 분석(分析))

  • Kim, Pil-Je;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
    • /
    • v.15 no.1
    • /
    • pp.37-45
    • /
    • 1996
  • Abstracts From the previous metabolic study of Pentachlorophenol(PCP), PCP was found to be exclusively transformed into ${\beta}-glucose$ conjugates of PCP in soybean and rice cell suspension cultures. In order to gather structural information of of the glucose conjugate, their aglycons and glycon have been analyzed by GC and GC/MS respectively, after thorough purification by chromatographic techniques. The glucose conjugates were effectively purified through a 1-butanol extraction followed by Silica gel TLC, Sephadex column chromatography and HPLC. Aglycons of the metabolites were identified as PCP, isomeric mixture of tetrachlorophenol, and tetrachlorocatechol and glycon were identified as glucose, suggesting that there are at least three kinds of glucose conjugates with different phenolic moieties. Under controlled conditions, the glucose conjugates were separated into three HPLC peaks which released respective aglycon upon a hydrolytic treatment. These results give valuable information on the structure of the glucose conjugates such that some PCP-driven chlorophenols, in addition to PCP, are also conjugated with glucose.

  • PDF

Isolation of GTP Binding Protein from Bovine Brain (소의 뇌로부터 GTP 결합단백질의 분리)

  • Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
    • /
    • v.10 no.2
    • /
    • pp.360-368
    • /
    • 1993
  • GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of $Go{\alpha}$ was revealed 39,000 dalton and $G{\beta}$ 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of $Go{\alpha}$ and $G{\beta}$.

  • PDF

Identification of interacting proteins of retinoid-related orphan nuclear receptor gamma in HepG2 cells

  • Huang, Ze-Min;Wu, Jun;Jia, Zheng-Cai;Tian, Yi;Tang, Jun;Tang, Yan;Wang, Ying;Wu, Yu-Zhang;Ni, Bing
    • BMB Reports
    • /
    • v.45 no.6
    • /
    • pp.331-336
    • /
    • 2012
  • The retinoid-related orphan nuclear receptor gamma ($ROR{\gamma}$) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the $ROR{\gamma}$ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro $ROR{\gamma}$-CTAP(SG), the nuclear localization of $ROR{\gamma}$-CTAP(SG) fusion protein was verified. Following isolation of $ROR{\gamma}$ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with $ROR{\gamma}$ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the $ROR{\gamma}$ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with $ROR{\gamma}$ protein in a complex format and function as co-activators in the $ROR{\gamma}$-mediated regulatory processes of HepG2 cells.

Inhibitory Effect of Elastase and Tyrosinase of Ginsenoside $F_1$ Isolated from Panax ginseng Leaves (인삼잎으로부터 분리된 진세노사이드 $F_1$의 Elastase 및 Tyrosinase 억제 효과)

  • Hong, Se Chul;Yoo, Nam Hee;Yoo, Ji Hyun;Lee, Kun Hee;Kim, Bo Ram;Lee, Ho Joo;Kim, Jong Min;Seong, Nak Sul;Pyo, Mi Kyung
    • Korean Journal of Pharmacognosy
    • /
    • v.44 no.1
    • /
    • pp.10-15
    • /
    • 2013
  • This study was carried to establish a simple isolation and purification method of ginsenoside $F_1$ from leaves of Panax ginseng and was to evaluate the inhibitory effect of purified ginsenoside $F_1$ on the activities of elastase and tyrosinase. The content of ginsenoside $F_1$ was 90-fold higher in leaves than in root of ginseng. Ginsenoside $F_1$ was isolated from EtOAc fraction between EtOAc and alkalized water of 80% EtOH extract after remove of hydrophobic components. The 50% inhibitory concentration ($IC_{50}$) of ginsenoside $F_1$ on elastase activity and tyrosinase activity was 1.07 mM and 1.81 mM, respectively. Especially, inhibitory effect of ginsenoside $F_1$ on tyrosinase activity was higher than that of arbutin ($IC_{50}$; 2.20 mM). These results indicate that ginsenoside $F_1$ have a potential for industrial cosmetic materials.

Developmental Changes of Serum IgA, IgG and IgM Concentrations in Broiler Chicks - III. Isolation of IgM and Developmental Changes of Serum IgM Levels (육계의 혈청중 면역글로부린(IgA, IgG, IgM)농도의 발육시기별 변화상 - III. IgM 분리 및 발육시기별 농도수준)

  • 김정우;이민호;김춘수;김상희;박근식
    • Korean Journal of Poultry Science
    • /
    • v.21 no.3
    • /
    • pp.175-182
    • /
    • 1994
  • An experiment was conducted to establish a large scale production method of anti-serum against chicken IgM and to profile the developmental changes of serum IgM levels during the feeding period(from hatching to 7 weeks of age) in broiler chicks. Blood samples were taken from Hubbard chicken at the age of hatching, three days of age, and weekly thereafter till to 7 weeks of age. The pure IgM was isolated from ammonium sulfate treated chicken serum by both sephadex G-200 and sepharose CL-6B chromatography. The breaking-through peak containing IgM appeared from the fraction 26 to 28. These fractions consisted mainly of IgM when tested by anti-chicken IgM(Nordic, Netherlands). Immunized with the heavy chain of this purified IgM, the rabbit immune sera(anti-chicken IgM) were formed a reaction only with the purified chicken IgM. The quantitative assay of serum IgM were carried by RID method. The optimal time for diffusion was 14 hours and the coefficient of determination($R^{2}$) for regression equation of standard curve was 0.992. It was observed that IgM concentrations were the highest at hatching(3.23 mg /mL), after that decreased gradually. From 2 to 5 weeks of age the levels unchanged(2.0 ~ 2.3mg /mL), and gradually decreased to 7 weeks of age(1.3 mg /mL).

  • PDF

Isolation and Properties of Amino Acid Antimetabolite from Streptomyces sp. 182-27 (Streptomyces sp. 182-27 균주가 생산하는 아미노산 대사길항물질의 정제와 특성)

  • 박부길
    • Microbiology and Biotechnology Letters
    • /
    • v.20 no.3
    • /
    • pp.335-343
    • /
    • 1992
  • A Streptomyces strain No. 182-27, which produced amino acid antimetabolite, was isolated from soil. During the course of screening for new amino acid antimetabolites from the culture broths of Actinomycetes, we found that the strain produced a substance active against Gram-positive bacteria and its activity was reversed by L-Ieucine on the synthetic minimal agar medium in the culture broth. The morphological and cultural characteristics serve to identify the producing organism strain 182-27 as the Streptomyces, although the species of this strain should be resolved in further studies. Fermentation was carried out in the synthetic medium at $28^{\circ}C$ for 78 hours. The fermentation yield reached about 2 mg per liter of the broth. Purification was done by ion exchange resin, active carbon, silica gel column chromatography and obtained 20 mg of pure active substance from the 20 $\ell$ culture broth. The 182-27 substance was obtained as white powder, mp 18SoC. From the physicochemical characteristics of the substance, it was amino acid like substance but unknown about its chemical structure. It is active against some Gram-positive bacteria and reversed by L-Ieucine.

  • PDF

Isolation and Characterization of Allelopathic Substances from Sorghum Stem (수수 줄기에 함유(含有)된 타감물질(他感物質)의 분리(分離) 및 특성(特性) 구명(究明))

  • Kim, S.Y.;De Datta, S.K.;Robles, R.P.;Kim, K.U.;Lee, S.C.;Shin, D.H.
    • Korean Journal of Weed Science
    • /
    • v.14 no.2
    • /
    • pp.156-162
    • /
    • 1994
  • To better understand the exact nature of the major toxic compound responsible for phytotoxicity of sorghum stem, the most toxic compound from the stem extract was isolated by rapid chromatography and subsequently purified by thin-layer chromatography(TLC) and high pressure liquid chromatography(HPLC). Of the eight fractions isolated by rapid chromatography, the fraction with solvent combinations of butanol (8) : acetic acid (1) : water (1) had the highest toxicity. Further separation of the fraction by TLC in a solvent mixture of butanol (24) : acetic acid (16.4) : water (7) : propanol (1) showed that the spot with an $R_f$ 0.71 had one major peak with retention time of 20.40 minutes. Upon subjecting gas chromatography and the HPLC fraction to the mass spectrometry, the toxic compound is probably one of the four compounds ; 1-methyl-1-(2-propynyl)-hydrazine, 1-aziridineethanol, 5-chloro-2-pentanone, and 2-(methylseleno)-ethanamine.

  • PDF