• Title/Summary/Keyword: HPLC screening

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

  • 이충환;백승화;김진희;김현아;이상명;이찬용;고영희
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.135-139
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    • 2003
  • During the screening for inhibitors of melanin biosynthesis from plant extract, Angelica polymorpha MAXIM which showed a high level of inhibition was selected. The inhibiting substances were purified form methanol extract of Angelica polymorpha MAXIM followed by silica gel column chromatography and HPLC. The inhibitors were identified as heraclenin, isosaxalin and heraclenol 3'-Me ether, by spectrescopic methods of ESI-MS, H-NMR, C-NMR, DEPT, HMQC and HMBC. These compounds did not have mushroom tyrosinase inhibitory activity, but showed a highly potent melanin biosynthesis inhibition zone in the plate culture of Streptomyces bikiniensis, a bacterium used as an indicator organism in this work. These compounds did not show any growth inhibition against S. bikiniensis at the same concentration of melanin biosynthesis test.

Neuraminidase Inhibitors from Mushroom Microphorus affinis

  • Kim, Kyung-Bum;Kim, Sang-In;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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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.

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

  • Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.2
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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.

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

  • Kim Hee Sook;Lee Eun Yeol
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.136-140
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    • 2005
  • Microbial cell-based UV spectrometric assay for the quantitative measurement of epoxide hydrolase activity was evaluated and optimized for the efficient screening of whole cell activity of novel epoxide hydrolase. Epoxide hydrolase activity was determined by measuring the increase of the oxidized product, benzaldehyde. The effects of the concentrations of phenyl-1,2-ethanediol, sodium metaperiodate and cells were optimized for epoxide hydrolase-catalyzed hydrolysis of styrene oxide. The relevant kinetic parameters of Km and $V_{max}$ for the hydrolysis of (R)-styrene oxide by Rhodotorula glutinis were determined from Lineweaver-Burk plot as 41.2 nmol/min$\cdot$mg dcw and 7.5 mM respectively, and coincided well with those from GC analysis.

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

  • Yun, Bong-Sik;Kim, Chang-Jin;Lee, In-Kyoung;Hiroyuki, Koshino;Yoo, Ick-Dong
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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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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DNA Toposiomerase I Inhibitor by Streptomyces sp. 7489 (방선균주 7489가 생산하는 DNA Topoisomerase I 저해제에 관한 연구)

  • Lee, Dong-Sun;Ha, Sang-Chul;Lee, Sang-Yong;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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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
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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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
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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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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    • v.15 no.3
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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.

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

  • 이선희;김미경;정미선;정용섭;엄태붕
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.18-22
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    • 1993
  • An assay system by using antibody was adopted to screen the endoinulinase producingfungi due to its high specificity toward endoinulinase, To determine whether the affinity-purified rabbit serum, which were generated against the purified endoinulinase, can react only with the endoinulinase, rocket immunoelectrophoresis was performed. The results showed that the serum specifically reacts with endoinulinase but not with exoinulinase and other proteins in the culture media. Using this polyclonal antibody, a strain from 62 fungal colonies was selected and it secreted an endoinulinase in the culture media to the amount comparable to that of Aspergillus ficuum A Tee 16882 known as a high endoinulinase producer.

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