• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.798-803
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• 1999

Prolyl endopeptidase [PEP; EC 3.4.21.26], a serine protease which is known to cleave peptide bonds on the carboxy side of a proline residue, plays an important role in the degradation of proline-containing neuropeptides that have been suggested to participate in learning and memory processes. An abnormal increase in the level of PEP, which can lead to generation of $A{\beta}$, is also suggested to be involved in Alzheimer's type senile dementia. In the course of screening PEP inhibitors from Basidiomycetes, the mushroom Polyozellus multiplex exhibited a high inhibitory activity against PEP. Two active compounds were isolated from the ethyl acetate soluble fraction by consecutive purification, using silica gel, Sephadex LH-20, and Lobar RP-18 chromatography. The chemical structures of these compounds were identified as thelephoric acid and 12-acety1-2,3,7,8-tetrahydroxy-[12H]-12-hydroxymethylbenzobis[I.2b,3.4b'] benzofuran-11-one (kynapcin-9) by spectral data including UV, IR, MS, HR-MS, $^1H-,{\;}^{13}C-$, and 2D-NMR. The $IC_{50}$ values of the thelephoric acid and kynapcin-9 were 0.157 ppm (446nM) and 0.087 ppm (212nM) and their inhibitor constants ($K_i$) were 0.73ppm ($2.09{\;}\mu\textrm{m}$) and 0.060 ppm (146 nM), respectively. Furthermore, they were non-competitive with a substrate in Dixon plots.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1359-1366
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• 2012

A strain of Streptomyces cavourensis subsp. cavourensis (coded as SY224) antagonistic to Colletotrichum gloeosporioides infecting pepper plants was isolated. SY224 produced lytic enzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase, and protease in respective assays. To examine for antifungal activity, the treatments amended with the nonsterilized supernatant resulted in the highest growth inhibition rate of about 92.9% and 87.4% at concentrations of 30% and 10%, respectively. However, the sterilized treatments (autoclaved or chloroform treated) gave a lowered but significant inhibitory effect of about 63.4% and 62.6% for the 10% supernatant concentration, and 75.2% and 74.8% for the of 30% supernatant concentration in the PDA agar medium, respectively, indicative of the role of a non-protein, heat stable compound on the overall effect. This antifungal compound, which inhibited spore germination and altered hyphal morphology, was extracted by EtOAc and purified by ODS, silica gel, Sephadex LH-20 column, and HPLC, where an active fraction was confirmed to be 2-furancarboxaldehyde by GS-CI MS techniques. These results suggested that SY224 had a high potential in the biocontrol of anthracnose in pepper, mainly due to a combined effect of lytic enzymes and a non-protein, heat-stable antifungal compound, 2-furancarboxaldehyde.

• Natural Product Sciences
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• v.13 no.2
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• pp.152-159
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• 2007

To demonstrate anti-hyperlipidemic activity of the 19${\alpha}$-hydroxyursane-type triterpenoid (19${\alpha}$-HUT)-rich fraction, this fraction was prepared from the extract of Rubus crataegifolius leaves. This fraction was found to have anti-hyperlipidemic effect in a high fat diet-induced rat model from the observation of reduction of abdominal fat pad weights, atherogenic index and hypercholesterolemia at 30 and 60 mg/kg (p.o.) The 19${\alpha}$-HUT fraction was subjected to SiO$_2$, ODS, and/or Sephadex LH-20 column chromatography to yield a new triterpenoid (1) called pomolic acid ester along with nine known triterpenoids which are all 19${\alpha}$-HUTs: euscaphic acid (2), tormentic acid (3), 23-hydroxytormentic acid (4), kaji-ichigoside F$_1$ (5), rosamultin (6), niga-ichigosides F$_1$ (7) and F$_2$ (8), suavissimoside F$_1$ (9) and coreanoside F$_1$ (10). The structure of compound 1 was established as 28-O-formyl-3,19-dihydroxyurs-12-en-28-oic acid on the basis of 2D-NMR spectroscopic data and mass spectrum. Compound 1 was isolated for the first time from natural sources.

• Journal of the Korean Wood Science and Technology
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• v.49 no.3
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• pp.267-273
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• 2021

Camellia nut shell was collected, dried at room temperature and ground to get fine powder. The powder was extracted three times with 95% EtOH, combined, evaporated, and then freeze dried. The crude powder was dissolved in H₂O and then sequentially fractionated with n-hexane, CH₂Cl₂, EtOAc and n-BuOH. A part of EtOAc fraction was chromatographed on a silica gel and on a Sephadex LH-20 columns using MeOH, aqueous MeOH, EtOAc-n-hexane and EtOH-n-hexane to isolate gallotannins. Three gallotannins, 1,2-di-O-galloyl-β-D-glucopyranoside (2), 1,2,6-tri-O-galloyl-β-D-glucopyranoside (3) and 1,2,3,6-tetra-O-galloyl-β-D-glucopyranoside (4), including gallic acid (1), were isolated and elucidated by NMR and Mass spectroscopies. Although nothing new, these gallotannins were first reported from the nut shell extractives of camellia tree (Camellia oleifera C. Abel). This study was to investigate the chemical constituents, especially hydrolysable tannins, of nut shell extractives of Camellia oleifera and to provide basic information for the future chemical utilization of this species.

• Mycobiology
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• v.47 no.2
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• pp.256-260
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• 2019

Neuraminidase (NA) cleaves the glycosidic bond linkages of sialic acids to release the mature virions from infected cells and has been an attractive therapeutic target for anti-influenza agents. In our ongoing investigation of NA inhibitors in mushroom extracts, we found that the extract the fruiting body of Glaziella splendens potently inhibited neuraminidase. The fruiting bodies of G. splendens were extracted and partitioned successively with hexane, ethyl acetate, and butanol. The ethyl acetate soluble-layer was subjected to silica gel and Sephadex LH-20 column chromatographies, and MPLC to obtain five compounds (1-5). Their structures were determined by spectroscopic methods. NA inhibitory activity of these compounds was evaluated using NAs from recombinant rvH1N1, H3N2, and H5N1 influenza A viruses. One compound (1) was elucidated as a new azaphilone derivative, and four compounds (2-5) were identified as entonaemin A, comazaphilone D, rubiginosin A, and entonaemin B, respectively. Compounds 3 and 4 showed considerable inhibitory activity against three types of neuraminidases with the $IC_{50}$ values of 30.9, 41.8, and $35.7{\mu}M$ for 3 and 46.5, 50.4, and $29.9{\mu}M$ for 4, respectively. This study reveals that the fruiting bodies of G. splendens possess azaphilone derivatives with the NA inhibitory activity. This is the first report on the isolation of neuraminidase inhibitors from the fruiting bodies of G. splendens.

• Journal of the Korean Wood Science and Technology
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• v.31 no.1
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• pp.32-40
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• 2003

Antimicrobial and antioxidative activities of heartwood extractives of domestic species were investigated to develop a natural fungicide or preservative. Four lignan derivatives and one taxane were isolated from heartwood of Taxus cuspidata which has been selected due to its high antioxidative activity among the tested species. The chemical structures were identified as : taxusin, isolariciresinol (4, 4', 9, 9'-tetrahydroxy-3', 5-dimethoxy-2, 7'-cyclolignan), lariciresinol (4, 4', 9-trihydroxy-3, 3'-dimethoxy-7, 9'-epoxylignan), taxiresinol (3, 4, 4', 9-tetrahydroxy-3'-methoxy-7, 9'-epoxylignan) and isotaxiresinol (3', 4, 4', 9, 9'-pentahydroxy- 5-methoxy-2, 7'-cyclolignan) on the basis of spectroscopic data and their chemical correlations. According to the results of free radical scavenging activity, isolariciresinol, lariciresinol and isotaxiresinol showed higher radical scavenging activity than those of 𝛼-tocopherol and butylated hydroxytoluene (BHT), the strongest natural and synthetic antioxidants. However, taxusin did not show any free radical scavenging activity. In this regard, it could inferred that high antioxidative activity of extractives of T. cuspidata was derived from isolariciresinol, lariciresinol and isotaxiresinol.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.4
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• pp.291-298
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• 1984

The extracted hagfish (Eptatretus burgeri) flesh lipid was separated into following fractions by column chromatography on Bio-beads SX-2 and Sephadex LH-20 prior to gab chromatographic analysis of their fatty acid compositions: polar lipid, triglyceride and free fatty acid. The major fatty acids of total lipid and triglyceride in hagfish were $C_{16:0},\;C_{16:1},\;and\;C_{18:1}$. The ratio of $C_{18:0}/C_{18:1}$ in the total lipid and triglyceride of hagfish was 0.1. The polar lipid of the hagfish muscle was mainly composed of phosphatidyl choline ($65.5\%$) and phosphatidyl ethanolamine ($28.0\%$). The triglyceride obtained was fractionated into four fractions by HPLC on the basis of partition numbers. Both the fatty acid composition and triglyceride composition on the basis of the total carbon number in the acyl chains of the triglyceride were analysed by the GLC. From the information obtained on triglyceride compositions based on the total carbon number by GLC and the partition number by HPLC and fatty acid composition by GLC, the combination of fatty acid in each triglycerides was estimated. A computer was used for estimation of the fatty acid combination in the triglyceride because hagfish lipid triglyceride was composed of various kinds of fatty acids. Fortyfour kinds of triglyceride were estimated. The major triglycerides in hagfish flesh lipid were found to those of ($1{\times}C_{16:0},\;2{\times}C_{18:1};\;13.5\%$), ($1{\times}C_{16:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;7.2\%$), ($1{\times}C_{16:1},\;2{\times}C_{18:1};\;5.4\%$), ($2{\times}C_{16:0},\;1{\times}C_{22:5};\;5.2\%$), ($1{\times}C_{14:0},\;2{\times}C_{18:1};\;4.5\%$), ($2{\times}C_{18:1},\;1{\times}C_{22:5};\;3.6\%$), ($1{\times}C_{14:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;2.7\%$) and ($1{\times}C_{14:0},\;1{\times}C_{16:0},\;1{\times}C_{18:2};\;2.2\%$).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.14 no.1
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• pp.1-6
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• 1981

In this study, sesame oil was chosen as the experimental sample and analysed for its triglyceride composition by high-performance liquid chromatography(HPLC) in combination with gas liquid chromatography(GLC). The triglycerides were separated from sesame oil by liquid chromatographies on Bio-Beads SX-2 and on Sephadex LH-20, and fractionated into five groups on the basis of their partition numbers by reverse phase HPLC on a column packed with $\mu-Bondapak$ C18 using methanol-chloroform mix-ture as a solvent. Each of these collected fractions gave one to three peaks in the GLC chromatograms according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analysed by GLC. From the results, it was found that the sesame oil consists with twenty one kinds of triglyceri-des, and the major triglycerides of sesame oil are those of $(2\;{\times}\;C18:1,\;C18:2\;;\;17.1\%),\;(C18:1,\;2{\times}C18:2\;;\;17.0\%),$ $(3\;{\times}\;C18:2\;;\;17.0\%),\;(3\;{\times}\;C18:1\;;\;10.9\%),$ $(3\;{\times}\;C18:2\;;\;9.6\%),\;(C16:0,\;C18:1,C18:2\;;\;7.9\%),$ $(C16:0,\;2\;{\times}\;C18:1\;;\;7.4\%),\;(C16:0,\;2\;{\times}\;C18:2\;;\;6.8\%),$ $(C18:0,\;C18:1,\;C18:2\;;\;3.1\%),\;(2\;{\times}\;C18:0,\;C18:2\;;\;1.5\%)$ $(C18:0,\;2\;{\times}\;C18:1\;;\;1.4\%),\;(C16:0,\;C18:0,\;C18:1\;;\;1.3\%),$ $(2\;{\times}\;C16:0,\;C18:1\;;\;1.2\%),\;and\;(C16:0,C18:0,\;C18:2\;;\;1.0\%)$.

• KSBB Journal
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• v.16 no.6
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• pp.592-602
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• 2001

This study was undertaken to investigate the antioxidative substance and activity of ethyl acetate extracted from Rumex crispus. Sample extracted follow in proper course of a solvent. Material refinement was carried out using silicagel column and Sephadex LH-20 column chromatography. Material sorting was carried by Gas Chromatography(GC/MS). 1,1-Diphenyl-2-Picrylhydrazyl(DPPH) free radical scavenging and enzyme activity were measured for antioxidative activity. as result of testing by DPPH free radical scavenging activity, Antioxidative activity was shown as the highest in the root, then leaf and stem in order. Ethyl acetate extraction of root part were 50% inhibitory concentration (IC50) Rumex activty(6.1 ug/mL). Rumex nipponicus(9.8 ug/ml) and Rumex acetoceae(31.5 ug/mL) in leaf part. The highest antioxidative activity of sample refined through silicagel column chromatography of Rumex crispus was appealed Fraction 5(IC50;3.57 ug/mL) in root and Fraction 6(IC50;85.9 ug/mL) in leaf. Fraction 5 in roof & Fraction 6 in leaf were refined using Sephadex LH-20 column chromatography. The highest antioxidative activity were appeared Fraction 4 (IC50;3.57 ug/mL) and Fraction 4 (IC50;18.41 ug/mL)in leaf. As for main phenol compounds 2,6-Dichloro-4-nitropnenol and 2-Isopropyl-5-methyl Phenol were identified in root and leaf, While 4-Vinyl-2- methoxy-phenol and 2,3-Dihydro- benzofuran were identifica ted only in leaf. Enzyme activity was shown low both in peroxidase(PDD) Non-activate(IU/mg protein)and in Superoxide dismutase(SOD) non-activate(IU/mg protein). 2,6-Dichloro-4-nitrophenol, 2-Isopropyl-5-methyl phenol, 4-Vinyl-2-methoxy-phenol were obtained in this experiment and these compounds are phenolic compounds which have OH group in the structure. With the result of this study these phenolic compounds which are extracted from Rumex crispus have high antioxidative effect. This antioxidative effect of Rumex crispus can be applied for chromo-preventive and antioxidative supplements which can be used for anti-allegy, aging, anti-tumor, aging and other oxidative disease for health promotion.

• Journal of Life Science
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• v.24 no.3
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• pp.234-241
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• 2014

To identify and isolate anticancer active compounds from Solanum nigrum, S. nigrum was extracted with MeOH and then fractionated with various organic solvents ($CH_2Cl_2$, EtOAc, n-BuOH, and $H_2O$). The cytotoxic effects of the MeOH extracts from S. nigrum and its organic solvent-soluble fractions were also tested in HT29 cells. All the MeOH extracts of S. nigrum and its organic-solvent extracts induced cytotoxicity in the HT29 cells. Among the extracts, $H_2O$ was the most effective. The $H_2O$ extract was purified further by repeated silica gel, Sephadex LH-20, Diaion HP- 20, and RP-18 column chromatography. An active anticancer compound, Des-N-26-methylene-dihydrotomatidine, was isolated with a molecular weight of 416 and a molecular formula of $C_{28}H_{48}O_2$. Analysis of the cytotoxic effects of Des-N-26-methylene-dihydrotomatidine on the HT29 cells compared to those of tomatine and tomatidine are similar in its structure, is higher than tomatidine above the 40 ${\mu}g/ml$ concentration, but lower than tomatine. This is the first study to describe the anticancer activity of Des-N-26-methylene-dihydrotomatidin, isolated from S. nigrum. Des-N-26- methylene-dihydrotomatidine seems to have potential as a natural bioactive compound.