• Natural Product Sciences
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• 제21권1호
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• pp.6-13
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• 2015

Phytochemical investigation of the aerial components of Ducrosia ismaelis Asch. led to the isolation of six known compounds, psoralen (1), isopsoralen (2), cnidioside A (3), (-)-syringaresinol-O-${\beta}$-D-glucopyranoside (4), (E)-plicatin B (5), trilinolein (6). The chemical structures of these compounds were elucidated from spectroscopic data and by comparison of these data with previously published results. The antioxidant, anti-osteoporotic and cardiovascular related activities of the isolated compounds were assessed using oxygen radical absorbance capacity (ORAC), reducing capacity, tartrate-resistant acid phosphatase (TRAP), and soluble epoxide hydrolase (sEH) inhibitory activity assays. Compounds (3-5) showed potent peroxyl radical-scavenging capacities with ORAC values of $11.06{\pm}0.39$, $7.98{\pm}0.10$, and $13.99{\pm}0.06$ Trolox equivalent (TE) at concentrations of $10{\mu}M$, respectively. Only compounds 4 and 5 was able to significantly reduce $Cu^{2+}$ ions, with a reduction value of $9.06{\pm}0.32$ and $4.61{\pm}0.00{\mu}M$ Trolox Equivalent (TE) at a concentration of $10{\mu}M$. Compound 5 at $10{\mu}M$ exhibited a potent inhibitory effect on osteoclastic TRAP activity with a TRAP value of $86.05{\pm}6.55%$ of the control. Compounds 1, 3 and 5 potently inhibited sEH activity with $IC_{50}$ values of 41.6 4.9, 16.0 1.1, and 49.0 $5.7{\mu}M$, respectively.

• 한국진공학회지
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• 제8권3B호
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• pp.262-268
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• 1999

Polycarbonate (PC) and Polymethylmethacrylate (PMMA) surface was modified by ion assisted reaction (IAR) technique to obtain the hydrophilic functional groups and improve the wettability. In conditions of ion assisted reaction, ion beam energy was changed from 500 to 1500eV, and ion dose and oxygen gas blown rate were fixed $1\times10^{16}$ ions/$\textrm{cm}^2$ and 4ml/min, respectively. Wetting angle of water on PC and PMMA surface modified by $Ar^+$ ion without blowing oxygen at 4ml/mon showed $5^{\circ}$ and $10^{\circ}$. Changes of wetting angle with oxygen gas and $Ar^+$ ion irradiation were explained by considering formation of hydrophilic group due to a reaction between irradiated polymer chain by energetic ion irradiation and blown oxygen gas. X-ray photoelectron spectroscopy analysis shows that hydrophilic groups such as -C-O, -(C=O)- and -(C=O)-O- are formed on the surface of polymer by chemical interaction. The polymer surface modification using ion assisted reaction only changed the surface physical properties and sept the bulk properties. In comparison with other modification methods, the surface modification by IAR treatment was chemically stable and enhanced the adhesion between metal and polymer surface. The applications of various kinds of polymer surface modification methods, metal and polymer surface. The applications of various kinds of polymer surface modification could be appled to the new materials about hydrophilic surface properties by IAR treatment. The adhesion between metal film and polymer measured by Scotch tape test whether the hydrophilic surfaces could improve the adhesion strength or not.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.722-727
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• 2005

Cytosolic sialidase (Neu2), a member of the sialidase family that is responsible for hydrolysis of sialic acid from the terminal position of sialoglycoconjugates, is poorly expressed in skeletal muscle and not detected in any other adult tissues. Thus, we isolated Neu2 cDNA using splicing by overlap extension (SOEing). In order to further characterize this enzyme, a His-tagged derivative was expressed in the bacterial expression system and purified by $Ni^{2+}$-affinity chromatography. A recombinant product of approximately 42 kDa had sialidase activity toward 4-methyl-umbelliferyl-$\alpha$-D-N-acetylneuraminic acid (4MU-NeuAc). The optimal pH and temperature of the recombinant Neu2 for 4MU-NeuAc was 6.0 and $37.5^{\circ}C$, respectively. The metal ions, such as $Cu^{2+}\;and\;Cd^{2+}$, showed strong inhibitory effect on the activity of the enzyme. The enzyme efficiently hydrolyzed the gangliosides GM3 and GD3 and had relatively low activities on ganglioside GD1a and GD1b, $\alpha$2-3 sialyllactose, and sialylated glycoproteins such as fetuin, transferrin, and orsomucoid, but had hardly any activities on $\alpha$2-6 sialyllactose and ganglioside GM1 and GM2. We concluded that the recombinant Neu2 has a sialidase activity toward glycoproteins as well as gangliosides.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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• pp.675-682
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• 2014

Approximately 50% of people in the world experience abdominal flatulence after the intake of foods containing galactosides such as lactose or soybean oligosaccharides. The galactoside hydrolyzing enzymes of ${\alpha}$- and ${\beta}$-galactosidases have been shown to reduce the levels of galactosides in both the food matrix and the human gastrointestinal tract. This study aimed to optimize the production of ${\alpha}$- and ${\beta}$-galactosidases of Bifidobacterium longum subsp. longum RD47 with a basal medium containing whey and corn steep liquor. The activities of both enzymes were determined after culturing at $37^{\circ}C$ at pH 6.0 for 30 h. The optimal production of ${\alpha}$- and ${\beta}$-galactosidases was obtained with soybean oligosaccharides as a carbon source and proteose peptone no. 3 as a nitrogen source. The optimum pH for both ${\alpha}$- and ${\beta}$-galactosidases was 6.0. The optimum temperatures were $35^{\circ}C$ for ${\alpha}$-galactosidase and $37^{\circ}C$ for ${\beta}$-galactosidase. They showed temperature stability up to $37^{\circ}C$. At a 1 mM concentration of metal ions, $CuSO_4$ inhibited the activities of ${\alpha}$- and ${\beta}$-galactosidases by 35% and 50%, respectively. On the basis of the results obtained in this study, B. longum RD47 may be used for the production of ${\alpha}$- and ${\beta}$-galactosidases, which may reduce the levels of flatulence factors.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.197-208
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• 2014

A novel protease gene from Bacillus gibsonii, aprBG, was cloned, expressed in B. subtilis, and characterized. High-level expression of aprBG was achieved in the recombinant strain when a junction was present between the promoter and the target gene. The purified recombinant enzyme exhibited similar N-terminal sequences and catalytic properties to the native enzyme, including high affinity and hydrolytic efficiency toward various substrates and a superior performance when exposed to various metal ions, surfactants, oxidants, and commercial detergents. AprBG was remarkably stable in 50% organic solvents and retained 100% activity and stability in 0-4 M NaCl, which is better than the characteristics of previously reported proteases. AprBG was most closely related to the high-alkaline proteases of the subtilisin family with a 57-68% identity. The secretion and maturation mechanism of AprBG was dependent on the enzyme activity, as analyzed by site-directed mutagenesis. Thus, when taken together, the results revealed that the halo-solvent-tolerant protease AprBG displays significant activity and stability under various extreme conditions, indicating its potential for use in many biotechnology applications.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.2090-2099
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• 2015

Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn²⁺, Ca²⁺, K²⁺, and Mg²⁺), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1359-1366
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• 2010

A search of the Streptomyces avermitilis genome reveals that its closest homologs are several O-methyltransferases. Among them, one gene (viz., saomt5) was cloned into the pET-15b expression vector by polymerase chain reaction using sequence-specific oligonucleotide primers. Biochemical characterization with the recombinant protein showed that SaOMT5 was S-adenosyl-L-methionine-dependent Omethyltransferase. Several compounds were tested as substrates of SaOMT5. As a result, SaOMT5 catalyzed O-methylation of flavonoids such as 6,7-dihydroxyflavone, 2',3'-dihydroxyflavone, 3',4'-dihydroxyflavone, quercetin, and 7,8-dihydroxyflavone, and phenolic compounds such as caffeic acid and caffeoyl Co-A. These reaction products were analyzed by TLC, HPLC, LC/MS, and NMR spectroscopy. In addition, SaOMT5 could convert phenolic compounds containing ortho-dihydroxy groups into O-methylated compounds, and 6,7-dihydroxyflavone was known to be the best substrate. SaOMT5 converted 6,7-dihydroxyflavone into 6-hydroxy-7-methoxyflavone and 7-hydroxy-6-methoxyflavone, and caffeic acid into ferulic acid and isoferulic acid, respectively. Moreover, SaOMT5 turned out to be a $Mg^{2+}$-dependent OMT, and the effect of $Mg^{2+}$ ion on its activity was five times greater than those of $Ca^{2+}$, $Fe^{2+}$, and $Cu^{2+}$ ions, EDTA, and metal-free medium.

• 한국광물학회지
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• 제9권1호
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• pp.1-6
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• 1996

Woolastonite from Susan occurs as intercalations in limestone beds of Lower Paleozoic Joseon Supergroup. It is a thermal metamorphic product of impure limestone. Electron microprobe analysis shows that it is considerably pure wollastonite. It has triclinic cell with a=7.932$\AA$, b=7.328$\AA$, c=7.069$\AA$, $\alpha$=89.995$^{\circ}$, $\beta$=$95.255^{\circ}$, and $ \Upsilon=103.367^{\circ}$.Dissolution behaviors of wollastonite have been studied conducting three different dissolution experiments; two different reactions with HC1 (one batch and one re-initialization experiment) and one traction with distilled water. In the batch type powder wollastonite-HCl reaction, pH of solution rapidly increases in the early stage and then its rate of increase slows down to reach plateau resulting in parabolic relationship with time. It is represented by the early rapid rise and fall in pH giving a sharp pH-edge and succeeding slow rise in the re-initialization experiment. The early rapid rise in pH is due to the rapid sorption of H- in solution to oxygens on the reactive surface of wollastonite and the fall in pH means that all reactive surface sites are occupied by H- ions and no more H- adsorption occurs. The slow rise in pH following the pH- edge is due to the dissolution of wollastonite as evidenced by the correlation of pH variation and cation concentration. Dissolution of powder wollastonite in HCl shows linear trend with time. Si is dissolved predominantly over Ca at a constant rate. Ca is dissolved predominantly in the very early stage. Dissolution rate of coarse-grained wollastonite fragments in distilled water is parabolic with times howing a rapid reaction in the early stage and a slow reaction in the advanced stage. The Ca/Si ratio in solution is high in the case of coarse-grained wollastonite fragment as compared with powder wollastonite. The coarse-grained wollastonite fragment-water (acid) reaction resulted in the solution with an elevated constant pH value (alkaline) giving an important significance on the environmental view point.

• 한국환경과학회지
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• 제16권12호
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• pp.1337-1343
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• 2007

The aim of this study was to isolate chicken feather-degrading bacteria with high keratinolytic activity and to investigate cultural conditions affecting keratinolytic enzyme production by a selected isolate. A chicken feather-degrading bacterial strain CH3 was isolated from poultry wastes. Isolate CH3 degraded whole chicken feather completely within 3 days. On the basis of phenotypical and 16S rDNA studies, isolate CH3 was identified as Bacillus thuringiensis CH3. This strain is the first B. thuringiensis described as a feather degrader. The bacterium grew with an optimum at pH 8.0 and $37^{\circ}C$, where maximum keratinolytic activity was also observed. The composition of optimal medium for keratinolytic enzyme production was feather 0.1%, sucrose 0.7%, casein 0.3%, $K_2HPO_4$ 0.03%, $KH_2PO_4$ 0.04%, $MgCl_2$ 0.01% and NaCl 0.05%, respectively. The keratinolytic enzyme had a pH and temperature optima 9.0 and $45^{\circ}C$, respectively. The keratinolytic activity was inhibited ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and metal ions like $Hg^{2+},\;Cu^{2+}\;and\;Zn^{2+}$. The enzyme activated by $Fe^{2+}$, dithiothreitol and 2-mercaptoethanol.

• 농업과학연구
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• 제44권2호
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• pp.261-271
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• 2017

The aim of this study was to investigate the amount of glucosinolates (GSLs) in kale sprouts (Brassica oleracea L. var. acephala) ('TBC') according to different concentrations of sulfur ions in sprout's nutrient solutions (0.0, 0.5, 1.0, and 2.0 mM) and to different light sources [Fluorescent lamp, Red, Blue, and Mix (R+B) LED]. Kale sprouts were cultivated in a growth chamber for 13 days in sulfur solutions. Kale sprouts were treated with fluorescent lamp and LED light sources for 5 days, from eight days after sowing to harvest. Amount of seven types of GSLs (progoitrin, sinigrin, 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, gluconasturtiin, and neoglucobrassicin) were measured in kale sprouts after harvest. The total GSL content was influenced by different sulfur solution concentration, and it was the highest at S 0.5 mM ($172.54{\mu}mol{\cdot}g^{-1}DW$) and the lowest at S 2.0 mM ($163.09{\mu}mol{\cdot}g^{-1}DW$). The GSL content was influenced by different light source, and it was the highest with Red LED ($159.23{\mu}mol{\cdot}g^{-1}DW$) and the lowest with Blue LED ($147.57{\mu}mol{\cdot}g^{-1}DW$). As the sulfur solution concentration increased under all light source, progoitrin and sinigrin contents tended to decrease while glucobrassicin content showed an upward tendency for all of the light sources. The content of glucobrassicin was higher than that of progitrin when treated with sulfur solutions for all LED light sources. Sinigrin, which has excellent anti-cancer effects, showed the highest rate (92.2%) among all the GSLs, under all of the light sources.