• 제목/요약/키워드: in-cell NMR

검색결과 393건 처리시간 0.027초

Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • 제22권1호
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Recently Isolated Bioactive Compounds from Korean Marine Sponges

  • Lim, Young-Ja;Kim, Jung-Sun;Chung J. Shim;Lee, Chong-O.;Im, Kwang-Sik;Jee H. Jung
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.89-94
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    • 1998
  • Marine sponges are recognized as a plentiful source of diverse biologically active secondary metabolites. Recently, we have initiated a research to discover antitumor constituents from the marine sponges collected from Korean Waters. Marine sponges collected from the South Sea of Korea were screened for several biological activities including such as brine shrimp lethality and cytotoxicity. Significant brine shrimp lethality was detected in the crude extract of a two-sponge association of Poecillastra sp. and Jaspis sp. A cross-section of this sample showed two layers of morphologically distinct sponges. The thin and dirty yellow outer layer was identified as Poecillastra sp. (Pachastrellidae), the surface of which was very rough. The light-grey inner layer was identified as Jaspis sp. (Jaspidae), the surface of which was smooth. This two-sponge association appears to be consistent as these sponges were always found in associated form regardless of collection site or collection period. Investigation of the bioactive constituents monitored by brine shrimp lethality assay led to the isolation of pectenotoxin II (PTX2) and psammaplin A as causative compounds for the brine shrimp lethality. $^1$H- and $\^$13/C-nmr signals of PTX2 was fully assigned utilizing TOCSY, HETCOR, Long-range HETCOR, and Homonuclear J-resolved 2D experiments. PTX2 displayed very potent and selective cytotoxicities in the 60 cell line panel antitumor assay at the NCI. PTX2 has progressed to acute toxicity determination and in vivo antitumor assay at the NCI (Table 1). However, significant in vitro antitumor activity of PTX2 can not be affirmed in the in vivo assay.

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Cytotoxic Isoflavanones from Uraria clarkei

  • Huang, Xiang-Zhong;Bai, Xi-Shan;Liang, Hui;Wang, Chao;Li, Wen-Juan;Guo, Jun-Ming;Jiang, Zhi-Yong
    • Bulletin of the Korean Chemical Society
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    • 제34권5호
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    • pp.1421-1424
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    • 2013
  • Two new isoflavanones, (3R) 5,7,3',4'-tetrahydroxy-2'-methoxyisoflavanone (1) and (3R) 5',8-di-(${\gamma}$,${\gamma}$-dimethylallyl)-2',5-dihydroxyl-4',7-dimethoxyl-isoflavanone (2), were isolated from Uraria clarkei, together with two known compounds dalbergioidin (3), 5,7-dihydroxy-2',4'-dimethoxyisoflavanone (4). The structures involving the absolute configuration of the new compounds were well elucidated by MS, IR, UV, CD, 1D and 2D NMR analyses. Cytotoxicity of the four compounds were assessed, results suggested that compound 2 possessed well cytotoxic activity, against the Hela, K562, and HL60 cell lines with $IC_{50}$ values of 28.0, 40.6 and $35.1{\mu}M$, respectively.

Melanin Biosynthesis Inhibition Effects of Ginsenoside Rb2 Isolated from Panax ginseng Berry

  • Lee, Dae Young;Jeong, Yong Tae;Jeong, Sang Chul;Lee, Mi Kyoung;Min, Jin Woo;Lee, Jae Won;Kim, Geum Soog;Lee, Seung Eun;Ahn, Young Sup;Kang, Hee Cheol;Kim, Jin Hee
    • Journal of Microbiology and Biotechnology
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    • 제25권12호
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    • pp.2011-2015
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    • 2015
  • Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng. Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, 1H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 μM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.

Fabrication of Micro Patterned Fibronectin for Studying Adhesion and Alignment Behavior of Human Dermal Fibroblasts

  • Lee, Seung-Jae;Son, Young-Sook;Kim, Chun-Ho;Choi, Man-Soo
    • Macromolecular Research
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    • 제15권4호
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    • pp.348-356
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    • 2007
  • The aim of this study was to fabricate a submicro-and micro-patterned fibronectin coated wafer for a cell culture, which allows the positions and dimensions of the attached cells to be controlled. A replica molding was made into silicon via a photomask in quartz, using E-beam lithography, and then fabricated a polydimethylsiloxane stamp using the designed silicon mold. Hexadecanethiol $[HS(CH_2){_{15}}CH_3]$, adsorbed on the raised plateau of the surface of polydimethylsiloxane stamp, was contact-printed to form self-assembled monolayers (SAMs) of hexadecanethiolate on the surface of an Au-coated glass wafer. In order to form another SAM for control of the surface wafer properties, a hydrophilic hexa (ethylene glycol) terminated alkanethiol $[HS(CH_2){_{11}}(OCH_2CH_2){_6}OH]$ was also synthesized. The structural changes were confirmed using UV and $^1H-NMR$ spectroscopies. A SAM terminated in the hexa(ethylene glycol) groups was subsequently formed on the bare gold remaining on the surface of the Aucoated glass wafer. In order to aid the attachment of cells, fibronectin was adsorbed onto the resulting wafer, with the pattern formed on the gold-coated wafer confirmed using immunofluorescence staining against fibronectin. Fibronectin was adsorbed only onto the SAMs terminated in the methyl groups of the substrate. The hexa (ethylene glycol)-terminated regions resisted the adsorption of protein. Human dermal fibroblasts (P=4), obtained from newborn foreskin, only attached to the fibronectin-coated, methyl-terminated hydrophobic regions of the patterned SAMs. N-HDFs were more actively adhered, and spread in a pattern spacing below $14{\mu}m$, rather than above $17{\mu}m$, could easily migrate on the substrate containing spacing of $10{\mu}m$ or less between the strip lines.

식물 대사체 연구의 진보 (Advances in Plant Metabolomics)

  • 김석원;정회일;유장렬
    • Journal of Plant Biotechnology
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    • 제33권3호
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    • pp.161-169
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    • 2006
  • 식물대사체학은 식물세포, 조직, 기관, 혹은 개체수준에서 주어진 시간과 조건에서 발견되는 모든 대사물질을 밝히고, 시간의 경과 혹은 조건의 변화에 따른 metabolic profiling의 변화를 연구하는 식물학 분야이다. 식물대사체학은 생물에 대한 전체론적 접근을 위한 가장 최근에 발달된 omics분야의 하나로서 일종의 시스템 생물학이다. 전체론적 접근과 이해를 위해서 대사체학은 metabolic profiling의 계량화학 혹은 다변량분석 방법을 자주 사용한다. 식물학분야에서 대사체학은 애기장대나 벼와 같은 모델식물에 tag를 도입하여 형질전환시킨 돌연변이체에 대해 DNA microarray와 함께 사용하여 유전자의 기능을 밝히는데 유용하게 사용된다. 본 총설에서는 식물대사체학의 기본 개념과 1H NMR 혹은 FTIR으로 얻은 metabolic profiling의 다변량분석에 대한 실용적인 사용법을 소개하고자 하였다.

Phenolic Compounds from the Fruit Body of Phellinus linteus Increase Alkaline Phosphatase (ALP) Activity of Human Osteoblast-like Cells

  • Lyu, Ha-Na;Lee, Dae-Young;Kim, Dong-Hyun;Yoo, Jong-Su;Lee, Min-Kyung;Kim, In-Ho;Baek, Nam-In
    • Food Science and Biotechnology
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    • 제17권6호
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    • pp.1214-1220
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    • 2008
  • Secondary metabolites from the fruit body of Phellinus linteus were evaluated for their proliferative effect on human osteoblast-like cells. 3-[4,5-Dimethylthiazole-2-y1]-2,5-diphenyl-tetraxolium bromide (MTT) assay and alkaline phosphatase (ALP) activity assay were used to assess the effect those isolates on the human osteoblast-like cell line (Saos-2). Activity-guided fractionation led to the isolation of ALP-activating phenolic compounds through the extraction of P. linteus, solvent partitioning, and repeated silica gel and octadecyl silica gel (ODS) column chromatographic separations. From the result of spectroscopic data including nuclear magnetic resonance (NMR), mass spectrometry (MS), and infrared spectroscopy (IR), the chemical structures of the compounds were determined as 4-(4-hydroxyphenyl)-3-buten-2-one(1), 2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde (2), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (3), 3,4-dihydroxybenzaldehyde (4), and protocatechuic acid methyl ester (5), respectively. This study reports the first isolation of compounds 1-3 and 5 from P. linteus. In addition, all phenolic compounds stimulated proliferation of the osteoblast-like cells and increased their ALP activity in a dose-dependent manner ($10^{-8}$ to $10^{-1}\;mg/mL$). The present data demonstrate that phenolic compounds in P. linteus stimulated mineralization in bone formation caused by osteoporosis. The bone-formation effect of P. linteus seems to be mediated, at least partly, by the stimulating effect of the phenolic compounds on the growth of osteoblasts.

떡윤노리나무로부터 분리된 Lyoniside의 함량분석과 항염증 효과 (Quantitative Analysis and Anti-inflammatory of Lyoniside from the Pourthiaea villosa var. brunnea)

  • 우경완;성태경;이혜미;장지훈;이기호;조현우;조정희;안병관
    • 생약학회지
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    • 제47권1호
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    • pp.12-17
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    • 2016
  • In this study, the purification of the MeOH extract from the stems and leaves of Pourthiaea villosa var. brunnea using column chromatography furnished a main compound, lyoniside. The structure was elucidated on the basis of $^1H$ and $^{13}C$ NMR spectroscopic data. Quantitative analysis of lyoniside was conducted by HPLC method and the highest content of lyoniside was found in 50% MeOH reflux extraction. To investigate the anti-inflammatory effect of the lyoniside, we measured nitric oxide and pro-inflammatory cytokines such as TNF-${\alpha}$, IL-$1{\beta}$, IL-6 levels in lipopolysaccharide-induced murine macrophage cell line RAW 264.7. As a results, lyoniside decreased the level of nitric oxide and IL-6 in concentration dose dependent manner in RAW 264.7 cells.

자란초에서 분리된 Luteolin 5-glucoside의 함량분석과 신경세포 보호 활성 (Quantitative Analysis of Luteolin 5-glucoside in Ajuga spectabilis and Their Neuroprotective Effects)

  • 우경완;심미옥;김아현;강병만;정호경;안병관;조정희;조현우
    • 생약학회지
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    • 제47권3호
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    • pp.211-216
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    • 2016
  • In the course of our continuing search for biologically active components from Korean medicinal plants, we isolated the main compound, luteolin 5-glucoside from aqueous fraction of Ajuga spectabilis. The structure was elucidated by the basis of $^1H$ and $^{13}C$ NMR and TOF ESI-MS data. Quantitative analysis of luteolin 5-glucoside was carried out on a XBridge C18 column ($S-5{\mu}m$, $4.6{\times}250mm$) with gradient elution composed of acetonitrile:water. The results exhibit that the average content of main compound in A. spectabilis were 0.048%. Oxidative stress plays a major role Alzheimer's disease (AD) and other neurodogenerative disease. AD is major health problem and there is currently no clinically accepted treatment to cure or stop its progression. Pretreatment with luteolin 5-glucoside markedly attenuated $H_2O_2$-induced cell viability loss in a dose-dependent manner. Luteolin 5-glucoside also inhibited the formation of intracellular reactive oxygen species in SH-SY5Y. The results suggest that luteolin 5-glucoside from A. spectabilis has protective effects against oxidative stress-induced cytotoxicity, which might be a potential therapeutic compound for treating and/or preventing neurodegenerative disease implicated with oxidative stress.

Multi-Function of a New Bioactive Secondary Metabolite Derived from Endophytic Fungus Colletotrichum acutatum of Angelica sinensis

  • Ramy S. Yehia
    • Journal of Microbiology and Biotechnology
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    • 제33권6호
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    • pp.806-822
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    • 2023
  • In the current study we assessed a new crystallized compound, 5-(1-hydroxybutyl)-4-methoxy-3-methyl-2H-pyran-2-one (C-HMMP), from the endophytic fungus Colletotrichum acutatum residing in the medicinal plant Angelica sinensis for its in vitro antimicrobial, antibiofilm, antioxidant, antimalarial, and anti-proliferative properties. The promising compound was identified as C-HMMP through antimicrobial-guided fraction. The structure of C-HMMP was unambiguously confirmed by 2D NMR and HIRS spectroscopic analysis. Antimicrobial property testing of C-HMMP showed it to be effective against a variety of pathogenic bacteria and fungi with MICs ranging from 3.9 to 31.25 ㎍/ml. The compound displayed excellent antibiofilm activity against C. albicans, S. aureus, and K. pneumonia. Furthermore, the antimalarial and radical scavenging activities of C-HMMP were clearly dosedependent, with IC50 values of 0.15 and 131.2 ㎍/ml. The anti-proliferative activity of C-HMMP against the HepG-2, HeLa, and MCF-7 cell lines in vitro was investigated by MTT assay, revealing notable anti-proliferative activity with IC50 values of 114.1, 90, and 133.6 ㎍/ml, respectively. Moreover, CHMMP successfully targets topoisomerase I and demonstrated beneficial anti-mutagenicity in the Ames test against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF). Finally, the compound inhibited the activity of α-glucosidase and α-amylase with IC50 values of 144.7 and 118.6 ㎍/ml, respectively. To the best of our knowledge, the identified compound C-HMMP was obtained for the first time from C. acutatum of A. sinensis, and this study demonstrated that C-HMMP has relevant biological significance and could provide better therapeutic targets against disease.