• 제목/요약/키워드: HPLC screening

검색결과 159건 처리시간 0.028초

궁궁이(Angelica polymorpha MAXIM)로부터 분리한 Coumarin 계열 화합물의 Melanin 생합성 억제 활성 (Melanin Biosynthesis Inhibitory Activities of Coumarins Isolated from Angelica polymorpha MAXIM)

  • 이충환;백승화;김진희;김현아;이상명;이찬용;고영희
    • 한국미생물·생명공학회지
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    • 제31권2호
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    • pp.135-139
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    • 2003
  • Melanin 생합성에 key enzyme인 tyrosinase 억제가 melanin색소 억제에 주된 요인으로 알려져 있지만, 생합성기작에는 여러 다른 요인들이 작용하기 때문에 일차적인 탐색 단계에서 tyrosinase 저해 활성뿐만 아니라 S. bikiniensis의 melanin생합성 억제 연구를 병행하여 천연물을 탐색하였다. 그 중 선택된 궁궁이 (Angelica poiymorpha MAXIM)에서 3가지 물질을 분리하였으며, 3가지 물질 모두가 100$\mu\textrm{g}$/$m\ell$ 이상 농도에서도 tyrosinase억제 활성이 나타나지 않는 반면, S. bikiniensis melanin 생합성 저해 효과를 보여 주었다. 동일 농도에서 S. bikinieffsis의 생장에는 영향을 미치지 않았다.

Neuraminidase Inhibitors from Mushroom Microphorus affinis

  • Kim, Kyung-Bum;Kim, Sang-In;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • 제13권5호
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    • pp.778-782
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    • 2003
  • In the course of screening anti-influenza agents from natural products, four neuraminidase inhibitors were isolated from the methanol extract of mushroom Microphorus affinis by purification using solvent partition, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC. The chemical structures of these compounds were identified as ${\alpha}-lupeol$, methyl linoleate, methyl palmitate, and methyl oleate by means of spectral data including GC-MS, $^1H-,\;and\;^{13}C-NMR\;with\;IC_{50}$ values of 5.65, 7.07, 7.12, and $7.52\;\mu\textrm{M}$, respectively. They did not inhibit other glycosidases such as glucosidase, mannosidase, and galactosidase, indicating that they were relatively specific inhibitors of neuraminidase. The relationship between the fatty acid structure and inhibitory activity was investigated. The result showed that, in the case of an aliphatic linear hydrocarbon skeleton, at least one carboxyl (presumably any carbonyl) moiety and sixteen carbons were the necessary requirements for potent inhibition, whereas saturated, unsaturated, free, and ester forms did not have any significant effect on the activity.

바지락(Ruditapes philippinarum) 아가미로부터 항균 펩타이드의 탐색 및 정제 (Screening and Purification of an Antimicrobial Peptide from the Gill of the Manila Clam Ruditapes philippinarum)

  • 서정길
    • 한국수산과학회지
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    • 제49권2호
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    • pp.137-145
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    • 2016
  • This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C18 reversed-phase and cation-exchange high-performance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.

미생물 세포 기반의 에폭사이드 가수분해효소 활성 측정을 위한 분광학적 분석법 최적화 (Optimization of Microbial Cell-Based Spectrometric Assay for the Analysis of Epoxide Hydrolase Activity)

  • 김희숙;이은열
    • 생명과학회지
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    • 제15권1호
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    • pp.136-140
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    • 2005
  • 다양한 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해 반응을 촉매하는 epoxide hydrolase 활성을 측정할 수 있는 미생물 세포 기반의 자외선 활성 분석법을 최적화하였다. 2.5 mg/ml의 세포 농도에서도 비교적 쉽게 흡광도 변화량을 인식할 수 있는 흡광도 범위인 0.5 이상을 얻을 수 있고, 반응 동력학 분석에도 응용할 수 있었다. 기존의 GC, HPLC 분석 법 보다 분석 시간을 줄일 수 있으며, 효소를 별도로 분리$\cdot$정제하지 않고 미생물 세포 자체의 epoxide hydrolase활성 분석이 가능하므로 상업적 특성이 우수한 epoxide hydrolase을 가진 미생물을 효율적으로 선별하는데 응용될 수 있을 것으로 기대된다.

Streptomyces sp. 3D3 균주가 생산하는 항고추역병성 항생물질 (An Antifungal Compound Against Phytophthora capsici Produced by Streptomyces sp. 3D3)

  • 윤봉식;김창진;이인경;히로유끼 고시노;유익동
    • 한국미생물·생명공학회지
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    • 제24권1호
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    • pp.77-81
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    • 1996
  • During the screening for the antifungal compounds against Phytophthora capsici causing phytophthora blight of red pepper, we isolated a strong active compound, bafilomycin $C_1$, produced by strain 3D3. The producing organism was identified as Streptomyces sp. based on taxonomic studies. The antifungal compound was purified from culture broth by HP-20 column chromatography, ethylacetate extraction, silica gel column chromatography and HPLC, and was identified as bafilomycin $C_1$ by color reaction, UV and $^{1}H$-NMR spectral data analysis. Bafilomycin $C_1$ showed strong antifungal activity against various phytopathogenic fungi.

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방선균주 7489가 생산하는 DNA Topoisomerase I 저해제에 관한 연구 (DNA Toposiomerase I Inhibitor by Streptomyces sp. 7489)

  • 이동선;하상철;이상용;김종국;홍순덕
    • 한국미생물·생명공학회지
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    • 제24권1호
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    • pp.101-104
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    • 1996
  • During the screening of inhibitor of DNA topoisomerase I from microbial secondary metabolites, Streptomyces melanosporofaciens 7489 which was capable of producing high level of inhibitor was selected from soil. The active compound (7489-1) was purified from the culture broth by solvent extraction, silica gel column chromatography and HPLC. The inhibitor was identified as dibutyl phthalate by spectroscopic methods of UV, $^{1}H$-NMR, $^{13}C$-NMR, DEPT and EI-MS. 7489-1 showed a strong inhibitory activity against topoisomerase I with 10 ${\mu}$M of $IC_{50}$ value.

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Screening of an antagonist of Pythium ultimum : Purification and characterization of an antibiotic effective to the oomycetes fungi

  • Yang, Jin-Ok;Park, Sang-Ho;Park, Dong-Jin;Kim, Chang-Jin
    • 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.135-135
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    • 1998
  • To find an antagonist of Pythium ultimum, the causal agent of damping-off, numerous actinomycete strains were screened for in vitro inhibiting mycelial growth of the target fungus and producing bioactive metabolites. A strain identified as Streptomyces sp. G60655 was isolated and used for further antagonistic efficacy. The degree of antagonism between the fungus and G60655 was affected by the medium used. Furthermore, the preinoculation of the antagonist was found to be necessary to exhibit the maximum efficacy of antagonsim against the fungus. From the culture broth, a bioactive metabolite was detected and purified by solvent extraction, silica gel chromatography and preparative HPLC. The FAB-MS spectrum of the active compound showed a molecular ion peak at m/z 1101 (M + H)$\^$+/, suggesting the molecular weight of 1100. The UV absorptions at 242 and 323 nm indicated the presence of aromatic functions. The structure of this compound was identified as echinomycin, a depsipeptide antibiotic by spectroscopic studies including various NMR measurements. Echinomycin was inactive against several soil born fungi, but inhibited the mycelial growth of P. ultimum and its related oomycetous fungi.

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GERI-BP001 Compounds, New Inhibitors of Acyl-CoA: Cholesterol Acyltransferase from Aspergillus fumigatus F37

  • Jeong, Tae-Sook;Kim, Sung-Uk;Son, Kwang-Hee;Kwon, Byoung-Mog;Kim, Young-Kook;Bok, Song-Hae
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.67-67
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    • 1995
  • Acyl-CoA:cholesterol acyltransferase (ACAT, EC 2.3.1.26) plays an important role in the control of intracellular free cholesterol content via its cholesterol esterifying activity. ACAT inhibitors are expected to be effective for treatment of atherosclerosis and hypercholesterolemia. In the course of a screening program for ACAT inhibitors from microbial sources, GERI-BP001 M, A, and B were isolated from the fermentation broth of a fungal strain. GERI-BP001 compounds were isolated from a culture broth of Aspergillus fumigatus F37 by acetone extraction, EtOAc extraction, SiO$_2$ column chromatography, and reverse phase HPLC. The structure of GERI-BP001 coumpounds were determined by $^1$H-NMR, $\^$l3/C-NMR, 2D-NMR, NOESY, and long range C-H COSY experiments. GERI-BP001 M, A, and B inhibit ACAT activity in an enzyme assay system using rat liver microsomes by 50% at concentrations of 75, 147, and 71 ${\mu}$M, respectively.

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Gentisyl Alcohol Inhibits Apoptosis by Suppressing Caspase Activity Induced by Etoposide

  • KIM JINHEE;KIM DONGHYUN;KIM MEEREE;KWON HOJEONG;OH TAEKWANG;LEE CHOONGHWAN
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.532-536
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    • 2005
  • In the course of our screening for small molecules to inhibit apoptosis of U937 human leukemia cells induced by etoposide ($10\;{\mu}g/ml$), Penicillium sp. F020150 with potent inhibitory activity was selected. The active compound was purified from ethyl acetate extract of the microorganism by Sephadex LH-20 column chromatography and HPLC, and was identified as gentisyl alcohol (2,5-dihydroxybenzyl alcohol) by spectroscopic methods. The compound inhibited caspase-3 induction with $IC_{50}$ value of $3.0\;{\mu}g/ml$ after 8 h of etoposide treatment. The expression levels of caspase-3 and PARP were dose-dependently inhibited by the compound, suggesting that gentisyl alcohol inhibits etoposide-induced apoptosis via downregulation of caspases.

항체를 이용한 Endoinulinase 생산 곰팡이의 검색 (Screening of the Endoinulinase-producing Fungi by Using Antibody)

  • 이선희;김미경;정미선;정용섭;엄태붕
    • 한국미생물·생명공학회지
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    • 제21권1호
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    • pp.18-22
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    • 1993
  • Oligofructo당 생산에 이용될 수있는 endoinulinase 분석방법은 그 균주가 endo- 및 exoinulinase를 함께 내는 경우, 일반적인 환원당 분석법으로는 정량하기가 어려워진다. 이 실험에서는 endoinulinase만을 선택적으로 정량하기 위한 하나의 방법으로써 항체 분석법을 이용하였다. Aspergillus niger ATCC 16882 조효소액을 CM-DEAE ion exchange chromatography, pI 2.5-5에서 preparative isoelectric focusing, HPLC gel filtration을 통해 순수하게 endoinulinase에 대한 항체를 얻었다.DEAE-ion exchange 및 protein A에서 정제된 이항체는 immunoassay 한 결과, exoinulinase 가 아닌 endoinulinase와 만 특이하게 반응하였고, immuno affinity chromatography 결과들은 배양액 중의 다른 단백질과 반응하지 않음이 확인되었다. 이눌린을 유일한 탄소원으로 한 배지에서 자란 1200여개의 야생균주들로부터 배양 특성이 우수한 균주를 1차로 선별하고 이 균주들의 endoinulinase 함량을 rocket immunoassay를 통하여 조사하였다. 이 중 1개의 균주는 Novozyme의 ATCC 1688와 비교할만한 정도의 endoinulinase를 배양액 중에 분비함을 확임할 수 있었다.

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