• Fisheries and Aquatic Sciences
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• 제17권2호
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• pp.215-222
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• 2014

To explore marine microorganisms with medical potential, we isolated and identified marine bacteria from floats, marine algae, animals, and sponges collected from Jeju Island, Korea. We isolated and identified 21 different strains from the marine samples by 16S rRNA analysis, cultured them in marine broth, and extracted them with ethyl acetate (EtOAc) to collect secondary metabolite fractions. Next, we evaluated their anti-oxidative and anti-inflammatory effects. Among the 21 strains, the secondary metabolite fraction of Bacillus badius had both strong antioxidant and anti-inflammatory activity, and thus was selected for further experiments. An antioxidant compound detected from the secondary metabolite fraction of B. badius was purified by preparative centrifugal partition chromatography (n-hexane:EtOAc:methanol:water, 4:6:4:6, v/v), and identified as diolmycin A2. Additionally, diolmycin A2 strongly inhibited nitric oxide production. Thus, we successfully identified a significant bioactive compound from B. badius among the bacterial strains collected from Jeju Island.

• 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.

• Fisheries and Aquatic Sciences
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• 제18권1호
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• pp.35-44
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• 2015

We previously identified Streptomyces griseus as an anti-cancer agent (Kim et al., 2014). In this study, we isolated compounds from S. griseus and evaluated their anticancer effect and toxicity in vitro and in vivo. Preparative centrifugal partition chromatography (CPC) was used to obtain three compounds, cyclo($_{\small{L}}$-[4-hydroxyprolinyl]-$_{\small{L}}$-leucine], cyclo($_{\small{L}}$-Phe-trans-4-hydroxy-$_{\small{L}}$-Pro) and phenethyl acetate (PA). We chose PA, which had the highest anticancer activity, as a target compound for further experiments. PA induced the formation of apoptotic bodies, DNA fragmentation, DNA accumulation in $G_0/G_1$ phase, and reactive oxygen species (ROS) formation. Furthermore, PA treatment increased Bax/Bcl-xL expression, activated caspase-3, and cleaved poly-ADP-ribose polymerase (PARP) in HL-60 cells. Simultaneous evaluation in vitro and in vivo, revealed that PA exhibited no toxicity in Vero cells and zebrafish embryos. We revealed, for the first time, that PA generates ROS, and that this ROS accumulation induced the Bcl signaling pathway.

• Food Science and Biotechnology
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• 제14권2호
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• pp.263-267
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• 2005

In order to isolate hyaluronidase (HAase) inhibitor from Astragali radix (AR), dried roots were extracted with ethanol, prior to sequential fractionations with n-hexane, chloroform, ethyl acetate, n-butanol, and aqueous fractions. The n-butanol soluble fraction was found to exhibit the most pronounced inhibitory effect (68%) on HAase, and the active components were separated using various chromatographic methods, including column chromatography and preparative HPLC. The active component was isolated from the n-butanol soluble fraction of AR and was structurally identified as calycosin-7-O-${\beta}$-D-glucopyranoside by LC-MS, IR, $^1H$ NMR, and $^{13}C$ NMR analysis. The $IC_{50}$ of calycosin-7-O-${\beta}$-D-glucopyranoside's HAase activity was found to be 3.7 mg/mL.

• Natural Product Sciences
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• 제15권2호
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• pp.106-109
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• 2009

Phytochemical investigations of the EtOAc-soluble fraction of the whole plants of Salvia plebeia using repeated column chromatography with preparative HPLC led to the isolation of a new phenylbutanone glucoside, 4-{4-0-[6-(4-hydroxybenzoyl)-0-${\beta}$-D-glucopyranosyl]-3-hydroxyphenyl}-butan-2-one (salviaplebeiaside, 1) along with two known phenolic compounds, rosmarinic acid methyl ester (2) and luteolin-7-0-${\beta}$-D-glucoside (3). The structures of these compounds were determined on the basis of spectroscopic methods including 1D-, 2D-NMR and MS spectrometry and comparison of spectroscopic data with those of values reported in the literatures.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.270-276
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• 1998

Extracellular ${\alpha}$-glucosidase was purified to homogeneity from moderately thermophilic Bacillus sp. DG0303. The thermostable ${\alpha}$-glucosidase was purified by ammonium sulfate fractionation, ion-exchange chromatography, preparative polyacrylamide gel electrophoresis (PAGE), and electroelution. The molecular weight of the enzyme was estimated to be 60 kDa by SDS-PAGE. The optimum temperature for the action of the enzyme was at $60^{\circ}C$. It had a half-life of 35 min at $60^{\circ}C$. The enzyme was stable at the pH range of 4.5~7.0 and had an optimum pH at 5.0. The enzyme preparation did not require any metal ion for activity. The thermostable ${\alpha}$-glucosidase hydrolyzed the ${\alpha}$-1,6-linkages in isomaltose, isomaltotriose, and panose, and had little or no activity with maltooligosaccharides and other polysaccharides. The $K_m$ (mM) for p-nitrophenyl-${\alpha}$-D-glucopyranoside (pNPG), panose, isomaltose, and isomaltotriose were 4.6, 4.7, 40.8, and 3.7 and the $V_{max}$(${\mu}mol{\cdot}min^-1$$mg^-1$) for those substrates were 5629, 1669, 3410, and 1827, respectively. The N-terminal amino acid sequence of the enzyme was MERVWWKKAV. Based on its substrate specificity and catalytic properties, the enzyme has been assigned to be an oligo-1,6-glucosidase.

• Natural Product Sciences
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• 제21권4호
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• pp.231-236
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• 2015

Two new azaphilone derivatives containing 1,3-dioxolane moiety, penidioxolanes A (1) and B (2), were isolated from marine-derived fungus Penicillium sp. KCB12C078, together with four known compounds (3-6) by chemical investigation. Compounds 1 - 6 were isolated by combination of silica gel, ODS column chromatography and preparative HPLC. Their structures were determined by analysis of spectroscopic data including 1D-, 2D-NMR, and MS techniques. The isolates were evaluated against cancer cell growth inhibition effects and antimicrobial activity.

• Food Science and Biotechnology
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• 제14권5호
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• pp.621-627
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• 2005

Halitosis is mainly caused by the presence of volatile sulfur-containing compounds (VSC's) produced by proteolytic periodontopathic bacteria in the oral cavity. Various mouth-rinses have been offered on the market as solutions to reduce halitosis. The aim of this study was to find a potent substance for the prevention of halitosis. The halitosis prevention substance (HPS) from cumin seed powder was purified by solvent extraction, silica gel column chromatography and preparative TLC to yield an oil phase (0.98%). Instrumental analysis such as FT-IR, $^1H$-NMR and $^{13}C$-NMR showed that HPS contained an -OH group, -HC=CH-, -COO-, and long chain acyl group. HPS was therefore determined to be 2-hydroxyethyl-${\beta}$-undecenate. HPS inhibited the growth of Fusobacterium nucleatum and Porphyromonas gingivalis, by 72.44% and 64.37% at $1{\times}10^{-2}\;M$, and by 99.85% and 91.62% at $5\;{\times}\;10^{-2}\;M$, respectively. It also inhibited the activity of L-methionine-${\alpha}$-deamino-${\gamma}$-mercaptomethane-lyase (METase), which was produced by oral microbes. Furthermore, the VSC production by oral microbes in the human mouth air decreased with increasing HPS concentration. These results suggested that HPS from cumin seed is an efficient halitosis prevention agent.

• Proceedings of the Korean Society of Applied Pharmacology
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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.

• Korean Journal of Microbiology
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• 제27권1호
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• pp.35-42
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• 1989

The mode of transglycosylation reaction observed during the action of low-molecular-weigh $\beta$-D-glucosidase ($\beta$-D-glucoside glucohydrolase, EC3.2.1.21) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H$-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [$\beta$-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [$\beta$-D glucopyranosyl-(1,4)-$\beta$-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [$\beta$-D-glucopyranosyl-(1,2)-D-glucopyranose], respectively.