• 제목/요약/키워드: prep-HPLC

검색결과 53건 처리시간 0.021초

Myxococcus stipitatus JW111이 생산하는 Polyene계 항암활성물질 (Cytotoxic Polyene Antibiotics from Myxococcus stipitatus JW111)

  • 안종웅
    • Applied Biological Chemistry
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    • 제45권2호
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    • pp.114-118
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    • 2002
  • 점액세균의 대사산물을 대상으로 새로운 항암활성물질을 탐색하는 과정에서 해양퇴적물에서 분리한 균주 JW111의 균체 추출액이 인체유래의 각종 암세포에 대해 강한 활성을 나타냄을 발견하고, 먼저 생산균주인 JW111이 Myxococcus stipitatus임을 확인한 후 그 대사산물을 분리, 정제하여 구조결정한 결과, Phenalamide $A_1$, $A_2E, $A_3$임을 밝혔다. 이들은 공시된 감수성 암세포에 대해 모두 우수한 활성을 나타내었으며, $IC_50$의 값은 $0.23{\sim}0.50{\mu}g/ml$이었다. 또한 이들은 Adriamycin의 내성 세포주에 대해서도 감수성 세포주와 동일한 활성을 나타내어 내성을 나타내지 않음이 확인되었다.

유청단백질의 분리 및 단백질 분해 효소에 의한 유청단백질의 가수분해 양상 (Isolation of whey protein and hydrolysis pattern of whey protein by proteolytic enzyme)

  • 렌친핸드;배형철;정석근;남명수
    • 농업과학연구
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    • 제39권4호
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    • pp.561-568
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    • 2012
  • The aim of this study was to introduce a simple method for isolation of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin from cow's milk, and peptides produced by enzymatic hydrolysis of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and ${\beta}$-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.

Phytosterols and Lignans from the Sesame Dregs of Sesamum indicum

  • Kim, Hye-Min;Lee, Jeong-Min;Park, Jun-Yeon;Lee, Sul-Lim;Han, Saem;Kim, Hyun-Young;Son, Dong-Wook;Choi, Sang-Yoon;Lee, Sang-Hyun P.
    • 한국자원식물학회지
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    • 제21권6호
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    • pp.420-426
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    • 2008
  • Phytochemical investigation of the sesame dregs of Sesamum indicum was conducted by open column and prep-HPLC chromatography. Two phytosterols (1 and 2) and two lignans (3 and 4) were isolated from the MeOH extracts of sesame dregs, and identified as ${\beta}$-sitosterol (1), daucosterol (2), sesamin (3), and sesamolin (4) by spectral analysis. Although these compounds were already isolated from sesame, it is important that they were still main phytochemical components in the sesame dregs.

천연물로부터 Quorum Sensing 저해제의 탐색 (Detection of a Quorum-Sensing Inhibitor from the Natural Products)

  • 김태우;차지영;이준승;민복기;백형석
    • 생명과학회지
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    • 제18권2호
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    • pp.206-212
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    • 2008
  • 인간이 서로 간의 의사소통을 위해 언어를 사용하듯이, 세균의 경우도 외부 환경 변화를 신속히 감지하여 서로 효과적으로 대응하기 위해서 주변 세포들과 소통할 수 있는 세균만의 독특한 화학적 언어를 사용하는 것으로 알려져 있다. 특히, 일정 세포 농도에 도달했을 때 자체적으로 생산된 화학적 신호를 통해 개체 수를 인지하고 그에 따라 특정 유전자의 발현을 동시에 조절하는 quorum sensing (QS) 기작은 다양한 세균 종들에서 광범위하게 존재한다. 본 연구는 다양한 천연물 추출물들을 대상으로 QS 저해 활성을 확인하였는데 QS 지시균주인 Agrobacterium tumefaciens NT1과 화학적으로 합성한 QS autoinducers을 사용한 bioassay를 수행하였다. 그 결과 양배추, 파, 양파의 추출물들에서 QS 저해 활성을 확인하였고, recycling preparative HPLC (prep-HPLC)를 통한 정제 과정을 통해, 83분 지점의 peak에 해당하는 성분들이 공통으로 QS 저해 활성을 가지고 있음을 확인하였다. 따라서 그 QS 저해 성분을 QSI-83으로 지정하고 thin layer chromatography (TLC)를 통해 P. syringae pv. tabaci의 autoinducers 합성을 저해하는 활성을 가지고 있음을 확인하였다. 또한 열에 대한 안정성과 세균 생장에서의 영향을 조사하였는데, 그 결과 QSI-83은 열에 안정하며 세균의 생장에는 영향을 끼치지 않는 물질임을 확인하였다. 따라서 우리는 천연물로부터 분리된 새로운 성분이 QS 저해제로서 이용될 수 있음을 제안한다.

Radiolabeling of 11C-sertraline by fast and easy loop method with [11C]CH3OTf

  • Lee, Hak Jeong;Jeong, Jae Min;Lee, Sang-Yoon;Ido, Tatsuo
    • 대한방사성의약품학회지
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    • 제3권1호
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    • pp.32-37
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    • 2017
  • Cis-(1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tertrahydro-N-methyl-1-naphthalenamine (sertraline) hydrochloride from among selective serotonin reuptake inhibitors (SSRIs) is a treatment of major depression. For the differential diagnosis by metabolizing serotonin in a patient with neurological disorders, the radiolabeled $^{11}C$-sertraline was developed for non-invasive positron emission tomography in living brain and use the evaluation of new drug for SSRIs. We release the results of a fast and easy radiolabeling method applied a one-step loop method with $[^{11}C]CH_3OTf$ for routine clinical applications of $^{11}C$-sertraline. 1 mg of a precursor for $^{11}C$-sertraline in 0.1 mL DMF and $5{\mu}L$ of 1N NaOH, were injected into the loop of semi-prep high-performance liquid chromatography (HPLC). $[^{11}C]CH_3OTf$ was passed through the loop at room temperature (RT). The $^{11}C$-sertraline was separated by the semi-preparative HPLC. $^{11}C$-sertraline was eluted at 28.0 min was collected and evaluated by analytical HPLC and mass spectrometer. The total radiolabeling efficiency of $^{11}C$-sertraline was $30.7{\pm}8.7%$. The specific activity was $64.8{\pm}51.4GBq/{\mu}mol$. The radiochemical and chemical purities were higher than 99%. The mass spectrum of the product showed m/z peaks at 307.1 (M+1), indicating the mass of sertraline. By the one-step loop method with $[^{11}C]CH_3OTf$, $^{11}C$-sertraline could be quickly and easily prepared for clinical application.

$^{11}$C 표지 자동합성장치에서 루프법을 이용한 ($^{11}$C)(R)-PK11195의 간편한 합성법 (A Convenient Radiolabeling of [$^{11}$C](R)-PK11195 Using Loop Method in Automatic Synthesis Module)

  • 이학정;정재민;이윤상;김형우;최재연;이동수;정준기;이명철
    • Nuclear Medicine and Molecular Imaging
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    • 제43권4호
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    • pp.337-343
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    • 2009
  • 목적: (R)-1-(2-Chlorophenyl)-N-methyl-N-1-(1-methyl-propyl)-3-isoquinoline carboxamide ((R)-PK11195)는 말초형 벤조디아제핀 수용체 (PBR)의 친화성이 높으며, 활성화된 소교세포(microglia)의 PBR에 선택적으로 결합하는 것으로 알려져 있다. 본 연구에서는 말초형 벤조디아제핀 수용체 (PBR)의 FET용 리간드인[$^{11}$C](R)-PK11195를, 절차가 빠르고 간단한 [$^{11}$C]CH$_3$I 자동합성장치 및 루프법을 도입해서 합성하였다. 대상 및 방법: 사이클로트론에서 $^{14}$N(p,a)$^{11}$C핵반응에 의하여 생산된 [$^{11}$C]Co$_2$를 0.2 M LiAlH$_4$/THF (0.2 mL)로 환원한 다음 HI (1 mL)과 반응하여 [$^{11}$C]CH$_3$I를 생산하였다. 반응용매인 NaH DMF complex (0.1 mL)와 녹인 전구물질 (R)-N-desmethyl-PK11195 (1 mg)을 녹인 DMSO (0.1mL)를 섞은 혼합액을 HPLC의 루프에 미리 주입하고 [$^{11}$C]CH$_3$I를 상온에서 5분 동안 질소가스로 불어준 뒤에 semi-preparative HPLC로 [$^{11}$C](R)-PK11195를 분리하였다. 결과: 전구물질과 [$^{11}$C]CH$_3$I의 [$^{11}$C]메틸화 반응에서의 표지효율은 71.8$\pm$8.5% 이었다. 분리 후 얻은 [$^{11}$C](R)-PK11195의 비방사능은 11.8$\pm$6.4 GBq/$\mu$mol이었으며, 방사화학적 순도는 99.2% 이상이었다. C-11 표지 후 얻어진 (R)-PK11195의 물질의 질량 분석은 m/z 353.1 (M+1)으로 물질구조를 확인 할 수 이었다. 결론: 뇌염증에 의한 활성화 소교세포의 영상화를 위한 PET용 방사성의약품인 [$^{11}$C](R)-PK11195를 반복적으로 생산해야 하는 임상 적용을 위해, 합성절차가 빠르고 간단한 [$^{11}$C]CH$_3$I 자동화 합성장치를 1차 루프 방법을 이용하여 $^{11}$C을 표지 할 수 있었다. 이 연구를 통해서 [$^{11}$C](R)PK11195 표지 할 때 반응 단계를 줄이고 반응을 실온에서 할 수 있어서 표지과정을 보다 단순화할 수 있었으며, NaH의 DMF현탁액을 사용함으로써 보다 안전하게 생산할 수 있게 되었다.

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

  • 권남우;김재연;조용범;황방연;김준구;우선희;이문순
    • 한국약용작물학회지
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    • 제27권6호
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    • pp.419-426
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    • 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.

Evaluation of Antioxidant Activities of Peptides Isolated from Korean Fermented Soybean Paste, Chungkukjang

  • Kim, Sun-Lim;Chi, Hee-Youn;Kim, Jung-Tae;Hur, On-Sook;Kim, Deog-Su;Suh, Sae-Jung;Kim, Hyun-Bok;Cheong, Ill-Min
    • 한국작물학회지
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    • 제56권4호
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    • pp.349-360
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    • 2011
  • The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.

An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • 제15권5호
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    • pp.1001-1010
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    • 2005
  • Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

꾸지 뽕나무로부터 분리한 flavonoid계 화합물의 암세포성장 저해 및 항산화 활성 (Tumor cell growth inhibition and antioxydative activity of flavonoids from the stem bark of Cudrania tricuspidata)

  • 이인경;송경식;김창진;김환묵;오구택;유익동
    • Applied Biological Chemistry
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    • 제37권2호
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    • pp.105-109
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    • 1994
  • 꾸지 뽕나무의 줄기껍질로부터 P388 tumor cell line에 대한 세포 성장 저해활성과 흰쥐 간 microsome 분획의 과산화 지질을 사용하여 $Fe^{++}/\;ascorbate$법으로 측정한 결과 세포 성장 저해활성과 항산화활성을 갖는 5개의 flavonoid계 화합물을 분리하였다. 분리된 화합물의 구조분석을 실시한결과 각종 spectral data와 보고된 문헌에 의하여 taxifolin, orobol, eriodictyol, dihydrokaempferol, steppogenin으로 각각 동정되었다. 이 화합물들의 항산화활성은 $IC_{50}$이 각각 6, 3, 3, >50, $10\;{\mu}g/ml$이었고 taxifolin을 제외한 P388 cell line에 대한 세포 성장저해는 각각 $IC_{50}$이 0.18, 3.3, 15, $6.2\;{\mu}g/ml$이었다. 한편 Escherichia coli BE 1186, Salmonella thyphimurium SL 1102, Staphylococcus aureus IFO 12732, Staphylococcus aureus R 209, Candida albicans에 대해서는 항균활성을 나타내지 않았다.

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