• Archives of Pharmacal Research
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• v.28 no.4
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• pp.395-399
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• 2005

From $70\%$ ethanol extract of the roots of Smilax bockii warb., seven flavonoids, kaempferol (1), $kaempferol-7-O-\beta-D-glucopyranoside$ (2), quercetin (3), isorhamnetin (4), (+)-dihydro­kaempferol (5), engeletin (6), isoengeletin (7), and $n-butyl-\beta-D-fructopyranoside$ (8), caffeic acid n-butyl ester (9) were isolated and identified by means of chemical and spectroscopic. Compounds 2, 4, and 6-9 were isolated for the first time from the roots of S. bockii and compounds 2, 8, and 9 were firstly isolated from the genus Smilax. In addition, using the SEAP (Secreted alkaline phosphatase) assay system, we investigated the in vitro anti-inflammatory activity of the $70\%$ ethanol extract of the roots of S. bockii, which showed moderate activity in inhibiting $TNF-\alpha-induced NF-{\kappa}B$ activation with an $IC_{50}$ value of $166.6 {\mu}g/mL$.

• Natural Product Sciences
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• v.20 no.3
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• pp.211-215
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• 2014

Opuntia humifusa, also called as Cheonnyuncho, is a cactus widely cultivated in southern regions of Korea. It has been known to have diverse biological activities, but most of the studies were performed with the MeOH extracts or solvent-partitioned fractions. Furthermore, the efforts to identify the responsible compounds for the biological activities are very limited. In this study, we tested the inhibitory effect of extracts and solvent-partitioned fractions of O. humifusa against LPS-induced nitric oxide (NO) production in Raw264.7 cells. The butanol fractions of O. humifusa efficiently inhibited the production of NO in Raw264.7 cells, but it was not due to the reduction of cell viability. Bioassay-guided isolation of butanol fractions of O. humifusa allowed the isolation of three flavonoids isorhamnetin 3-O-${\beta}$-$\small{D}$-galactosyl-4'-O-${\beta}$-$\small{D}$-glucoside (1), isorhamnetin 3,4'-di-O-${\beta}$-$\small{D}$-glucoside (2) and isorhamnetin 3-O-${\beta}$-$\small{D}$-(6-O-${\alpha}$-$\small{L}$-rhamnosyl)glucoside (3), and one lignan syringaresinol O-${\beta}$-$\small{D}$-glucopyranoside (4). Among them, isorhamnetin 3-O-${\beta}$-$\small{D}$-galactosyl-4'-O-${\beta}$-$\small{D}$-glucoside (1) and isorhamnetin 3,4'-di-O-${\beta}$-$\small{D}$-glucoside (2) exhibited the moderate inhibitory effects against LPS-induced NO production. This is the first time to report anti-inflammatory effects of these compounds.

• The Korean Journal of Physiology
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• v.30 no.2
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• pp.257-267
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• 1996

Diabetes mellitus is associated with a wide range of pathophysiologic changes in the kidney. This study was designed to examine the mechanisms by which glucose modulates the expression of polarized membrane transport functions in primary cultured rabbit renal proximal tubule cells. Results are as follows: The rate of 30 minute $Rb^{+}$ uptake was significantly higher($137.76{\pm}5.40%$) in primary renal tubular cell cultures treated with 20 mM glucose than that of 5 mM glucose. Not the level of mRNA for the ${\alpha}$ subunit of Na, K-ATPase but that of ${\beta}$ subunit was elevated in primary cultures treated with high glucose. The initial rate of methyl-${\alpha}$-D-glucopyranoside(${\alpha}$-MG) uptake was significantly lower($71.91{\pm}3.02%$) in monolayers treated with 20 mM glucose than that of 5 mM glucose. There was a tendency of an increase in phlorizin binding site in cells treated with 5 mM glucose. However, 3-O-methyl-D-glucose(3-O-MG) uptake was not affected by glucose concentration in culture media. TPA inhibited $Rb^{+}$ uptake by $63.61{\pm}1.94\;and\;45.80{\pm}1.36%$ and ${\alpha}$-MG uptake by $48.54{\pm}3.69\;and\;41.87{\pm}6.70%$ in the cells treated with 5 and 20 mM glucose, respectively. Also TPA inhibited mRNA expression of Na/glucose cotransporter in cells grown in 5mM glucose medium. cAMP significantly stimulated ${\alpha}$-MG uptake by $114.65{\pm}5.70%$ in cells treated with 5mM glucose, while it did not affect ${\alpha}$-MG uptake in cell treated with 20 mM glucose. However, cAMP inhibited $Rb^{+}$ uptake by $76.69{\pm}4.16\;and\;66.87{\pm}2.41%$ in cells treated with 5 and 20 mM glucose, respectively. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Na/glucose cotransport system is inhibited. High glucose may in part affect the activity of the Na,K-ATPase and the Na/glucose cotransport system by controlling the protein kinase C and/or A signal transduction pathway in primary cultured renal proximal tubule cells.

• Korean Journal of Plant Tissue Culture
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• v.27 no.5
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• pp.415-421
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• 2000

The disaccharide trehalose ($\alpha$-D-glucopyranosyl-$\alpha$-D-glucopyranoside) is found in variety of organ-isms that are able to withstand almost complete desiccation. In order to identify the function of trehalose in plants, we isolated Arabidopsis trehalase (AtTRE) gene that encodes the enzyme able to hydrolyze trehalose to glucose, and trehalose-6-phosphate synthase isolog, TPS3 gene by RT-PCR. The AtTRE had the substrate specificity to hydrolyze only trehalose, and a broad pH range of enzyme activity. The AtTRE promoter/GUS reporter gene was expressed in cotyledons, mature leaf tissues including guard cells, and developing siliques. The GUS expression driven by AtTPS3 promoter was significant in root tissues, and the level of GUS activity was much higher than that of the pBll 21 control seedlings. The knockout of AtTPS3 gene in Arabidopsis resulted in the retarded root development, whereas the overexpression of AtTPS3 increased the root elongation in the presence of sucrose in MS medium. Possible functions of AtTRE and AtTPS3 in plant will be discussed. In addition, ectopic expression of yeast TPS1 driven by the inducible promoters in tobacco and potato conferred the plants on the drought and freezing tolerances.

• Journal of Dairy Science and Biotechnology
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• v.39 no.1
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• pp.9-19
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• 2021

Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.535-540
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• 2012

${\beta}$-D-Xylosidase (E.C. 3.2.1.37) from Bifidobacterium breve K-110, which hydrolyzes ginsenoside Ra1 to ginsenoside Rb2, was cloned and expressed in Escherichia coli. The ($His_6$)-tagged recombinant enzyme, designated as XlyBK-110, was efficiently purified using $Ni^{2+}$-affinity chromatography (109.9-fold, 84% yield). The molecular mass of XylBK-100 was found to be 55.7 kDa by SDS-PAGE. Its sequence revealed a 1,347 bp open reading frame (ORF) encoding a protein containing 448 amino acids, which showed 82% identity (DNA) to the previously reported glycosyl hydrolase family 30 of Bifidobacterium adolescentis ATCC 15703. The $K_m$ and $V_{max}$ values toward p-nitrophenyl-${\beta}$-D-xylopyranoside (pNPX) were 1.45mM and 10.75 ${\mu}mol/min/mg$, respectively. This enzyme had pH and temperature optima at 6.0 and $45^{\circ}C$, respectively. XylBK-110 acted to the greatest extent on xyloglucosyl kakkalide, followed by pNPX and ginsenoside Ra1, but did not act on p-nitrophenyl-${\alpha}$-L-arabinofuranoside, p-nitrophenyl-${\beta}$-D-glucopyranoside, or p-nitrophenyl-${\beta}$-D-fucopyranoside. In conclusion, this is the first report on the cloning and expression of ${\beta}$-D-xylosidase-hydrolyzing ginsenoside Ra1 and kakkalide from human intestinal microflora.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.26-32
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• 2013

Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

• BMB Reports
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• v.29 no.2
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• pp.163-168
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• 1996

Glycosylated polyphenol oxidase was purified from potato tuber using ammonium sulfate fractionation, Sephadex G-100, and concanavalin A Sepharose column chromatography. Two or three types of polyphenol oxidase were separated on concanavalin A Sepharose. Type I and II polyphenol oxidases did not bind to concanavalin A Sepharose. Type I seemed to be an aggregated form of polyphenol oxidase. Type III polyphenol oxidase, which is presumed to be glycosylated because it was bound to concanavalin A Sepharose and eluted with $\alpha$-D-methyl glucopyranoside, was further purified by chromatography on Econo-Pac Q and Superose 12. Glycosylated polyphenol oxidase was purified 130-fold from the dissolved ammonium sulfate pellet resulting in about $6\;{\mu}g$ of the enzyme from 100 g of potato tuber periderm. The molecular weight of the glycosylated enzyme determined by SDS-polyacrylamide gel electrophoresis was about 64,000. Optimum temperature and pH of both II and type III potato polyphenol oxidases were $20^{\circ}C$ and pH 7.0, respectively. Glycosylated form of polyphenol oxidase (type III) preferred catechol to catechin as a substrate, whereas type II enzyme showed the reverse substrate preference.

• Natural Product Sciences
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• v.17 no.3
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• pp.183-188
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• 2011

A bisdesmosidic cholestane glycoside was isolated from the rhizomes of Polygonatum sibiricum and the structure was elucidated by spectroscopic methods and acid hydrolysis as (22S)-cholest-5-ene-$1{\beta}$,$3{\beta}$,$16{\beta}$,22-tetrol 1-O-${\alpha}$-L-rhamnopyranosyl 16-O-${\beta}$-D-glucopyranoside. This compound exhibited weak cytotoxic activity with the $IC_{50}$ value, $63.6\;{\mu}M$ in human MCF-7 breast cancer line, whereas it failed to show agonistic activity at $100{\mu}M$ in TGR5 assay with Chinese hamster ovary (CHO) cells. This is the first report of a bisdesmosidic cholestane glycoside from Polygonatum species and the full assignments of $^1H$, $^{13}C$ NMR by HMBC, TOCSY and NOESY experiments were provided.

• Natural Product Sciences
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• v.17 no.2
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• pp.90-94
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• 2011

An in vitro bioassay-guide revealed that the methanol (MeOH) extract of the whole plant of Patrinia saniculaefolia (Valerianaceae) showed cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) dual inhibitory activity by assessing their effects on the production of prostaglandin $D_2$ ($PGD_2$) and leukotriene $C_4$ ($LTC_4$) in mouse bone marrow-derived mast cells (BMMCs). Phytochemical study of the MeOH extract of this plant led to the isolation of twelve compounds; ${\beta}$-farnesene (1), squalene (2), nardostachin (3), patridoid I (4), patridoid II (5), patridoid II-A (6), oleanolic acid (7), oleanonic acid (8), 23-hydroxyursolic acid (9), oleanolic acid 3-O-${\alpha}$-L-arabinopyranoside (10), oleanolic acid 3-O-${\beta}$-D-glucopyranoside (11), oleanolic acid 3-O-[${\beta}$-D-xylopyranosyl-(1${\rightarrow}$3)-${\beta}$-D-(6-O-butyl)glucuronopyranoside] (12). Among the compounds, 4 and 5 strongly inhibited both the COX-2-dependent $PGD_2$ generation with $IC_{50}$ values of 8.7 and 13.6 ${\mu}M$, respectively, and the generation of $LTC_4$ in the 5-LOX dependent phase with $IC_{50}$ values of 41.7 and 46.9 ${\mu}M$, respectively, which suggest that the anti-inflammatory activity of P. saniculaefolia might occur in part via the inhibition of both $PGD_2$ and $LTC_4$ generation by 4 and 5.