• Korean Journal of Food Science and Technology
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• v.20 no.6
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• pp.780-786
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• 1988

In order to investigate the most efficient and rapid method for the purification of enterotoxin A from Staphylococcus aureus M 7/1, various methods such as ion-exchange chromatography on Amberlite, and CM-cellulose. gel filtration on Sephadex G-50, 75, 100 and Sephacryl, and fast protein liquid chromatography (FPLC) were applied and compared in terms of purity and speed. Although ion-exchange chromatography on Amberlite resin was good enough to remove other materials in culture medium from enterotoxin, and convenient, and fast method, the purity of this method was less than 70%. However. carboxymethyl ion-exchange column showed to be better purity than that of Amberlite method. The yields of these two methods were about 70% and 75%, respectively. When gel filtration methods on Sephadex G-50, 75, 100 and Sephacryl were applied, the purity was about 90%. Fast protein liquid chromatography was found to be the most efficient method in terms of purity (97%) and speed. The combined method, gel filtration after CM-cellulose column (stepwise elution) treatment can be also used as a efficient method particularly for the purification of large volume of sample.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.6-12
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• 1998

To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% $(NH_4)_2SO_4$ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and $60^{\circ}C$, respectively. The enzyme was stable in pH 7.0-12.0 at $50^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $Pb^{2+}$, whereas it was activited by $Na^+$, $Mg^{2+}$ and $Mn^{2+}$. The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were $29.33\;{\mu}mole/L$ and $5.13\;{\mu}g/min$, respectively.

• Applied Biological Chemistry
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• v.46 no.3
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• pp.251-256
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• 2003

The polyphenol fractions of Korean ginseng were purified using Sephadex LH-20, MCI gel, Bondapak $C_{18}$ TLC, and HPLC from the 60% acetone soluble fraction. Fraction I showed 48.16%, 79.71% and 43.55% inhibition at 150 ppm against lipid oxidation in the presence of copper ion, superoxide and hydrogen peroxidation. Electron donating abilities of fraction II showed 35.17% inhibition at 200 ppm. Fraction III showed 48.49% and 25% inhibition at 150 ppm against lipid oxidation in the presence of ferrous ion and hydroxy radical ion. The phospholipase $A_2$ inhibitory effect of fraction III was 48.9% at the concentration of $60\;{\mu}g/ml$. The cytotoxic effects of fraction II was the highest (73.29% at 0.25 mg/ml) among the tested polyphenol fractions.

• The Journal of Korean Academy of Prosthodontics
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• v.47 no.4
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• pp.376-384
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• 2009

Purpose: The purpose of this study was to evaluate the response of osteoblast-like cells to Ca-P coated surface obtained via Ion beam-assisted deposition (IBAD) method and Sol-Gel process on anodized surface by cellular proliferation and differentiation. Material and methods: The surface of a commercially pure titanium (Grade IV) discs with dimension of 10mm diameter and 2 mm thickness was modified by anodic oxidation under a constant voltage of 300 V. The experimental groups were coated with Ca-P by the IBAD method and Sol-Gel process on anodized surface. The surface roughness (Ra) of specimens was measured by optical interferometer and each surface was examined by SEM. To evaluate cell response, MG63 cells were cultured and cell proliferation, ALP activity and the ability of cell differentiation were examined. Also, cell morphology was examined by SEM. The significant of each group was verified by Kruskal-Wallis Test ($\alpha$=.05). Results: The Ra value of Ca-P coated surface by IBAD method was significantly higher than Ca-P coated surface by Sol-gel process (P < .05). The level of cell proliferation and ALP activity was higher in Ca-P coated surface by IBAD method (P<.05). The expression of ALP showed higher level expression in Ca-P coated surface by IBAD method. Cells grown on Ca-P coated surface by IBAD method were uniformly distributed and developed a very close layer. Conclusion: These experiments showed better performances of Ca-P coated surface by IBAD method with respect to Ca-P coated surface by Sol-gel process. Ca-P coated surface by IBAD method appear to give rise more mature osteoblast characteristics and might result in increased bone growth and bone-implant contact.

• Journal of Radiation Protection and Research
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• v.8 no.1
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• pp.15-25
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• 1983

The sequential separation of Ru-103, Cs-137 and Ce-144 was carried out by organic cation exchanger, Amberite CG-120, and inorganic ion exchangers, silica gel and montmorillonite. The optimum conditions of Ru-103, Cs-137 and Ce-144 on Amberite CG-120 are 0.01M-, 0.01M- and 0.1IM- hydrochloric acid for the adsorption, and 3M-, 3M- and 5M-hydrochloric acid for the desorption, respectively. The optimum conditions of Ru-103, Cs-137 and Ce-144 on silica gel are pH 8, pH 8 and pH 8 for the adsorption. and 3M-, 1M- and 1M-hydrochloric acid for the desorption. respectively. The optimum conditions of Ru-103, Cs-137 and Ce-144 on montmorillonite are pH 8, 0.01M-hydrochloric acid and pH 4 for the adsorption, and 1M-, 5M- and 3M-hydrochloric acid for the desorption. respectively. The adsorption which occurs at lower ionic strength and the differences in desorption ionic strength are utilized for the separation of tracer mixture in continuous experiments. The individual separation of Ru-103, Cs-137 and Ce-144 can be carried out more efficiently with montmorillonite than with silica gel and Amberite CG-120.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.574-582
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• 1992

Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

• Corrosion Science and Technology
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• v.14 no.2
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• pp.64-75
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• 2015

Single $TiO_2$ coating prepared by sol-gel process usually experiences cracks in coating layer. In order to prevent cracks, an inorganic-organic hybrid $TiO_2-SiO_2$ coating was synthesized by combining precursors with an organic functional group. Five different coatings with various ratios of (1:8, 1:4, 1:1, 1:0.25 and 1:0.125) titanium alkoxide (TBOT, Tetrabutylorthotitanate) to organo-alkoxysilane (MAPTS, ${\gamma}$-Methacryloxy propyltrimethoxysilane) on carbon steel substrate were made by sol-gel dip coating. The prepared coatings were analyzed to study the coating properties (surface crack, thickness, composition) by scanning electron microscope (SEM), focused ion beam (FIB), and Fourier transform infrared spectroscopy (FT-IR). Potentiodynamic polarization tests and electrochemical impedance spectroscopy (EIS) were also performed to evaluate the corrosion characteristics of the coatings. Crack free $TiO_2-SiO_2$ hybrid coatings were prepared with the optimization of the ratio of TBOT to MAPTS. The corrosion rates were significantly decreased in the coatings for the optimized precursor ratio without cracks.

• Journal of the Korean Electrochemical Society
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• v.6 no.2
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• pp.98-102
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• 2003

Urethane acrylate oligomer was synthesized and used in a gel polymer electrolyte (GPE) and then its electrochemical performances were evaluated. $LiCoO_2/GPE/graphite$ cells were prepared and their performances depending on discharge currents and temperatures were evaluated. The precursor containing $5 vol\%$ curable mixture had a low viscosity relatively. ionic conductivity of the gel polymer electrolyte at room temperature and $-20^{\circ}C$ was ca. $5.9\times10^{-3}S{\cdot}cm^{-1}\;and\;1.4times10^{-3}S{\cdot}cm^{-1}$, respectively. GPE showed good electrochemical stability up to potential of 4.5V vs. RLi/Li^+.\;LiCoO_2/GPE/graphite$ cell showed a good high-rate and low-temperature performance.

• Korean Journal of Microbiology
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• v.26 no.2
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• pp.73-81
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• 1988

$\alpha$-Amylase (EC.3.2.1.1) of Neurospora crassa (ATCC9279) was cloned in E. coli HB101 using shotgun method, and the enzymes isolated from both N. crassa and E. coli were compared. Chromosomal DNA isolated from the spores of N. crassa was partially digested with PstI restriction endonuclease and rejoined to pBR322 which had been digested with the same enzyme. The resulting recombinant DNA were introduced into E. coli HB101 which had competancy by treating with $CaCl_{2}$. As the result, about 8000 colonies which showed tetracycline resistance were selected and two of the colonies which had 13.5Kb recombinant plasmid exhibit starch degrading activity on starch-containing plate when treated with D-cycloserine. $\alpha$-Amylases from both N.crassa and E. coli were isolated by using ammonium sulfate precipitation, DEAE-cellulose ion exchange column chromatography and Bio-Gel P150 gel foltration column. As the result, about 81.3 fold and 5.6 fold purifications in specific activities were obtained respectively, and specific activities of the gel filtrates were 6.1u/mg and 85u/mg respectively. The properties of both enzymes were compared and they showed quite the similar patterns in optimal temperature, optimal pH and had same molecular weight about 100,000 daltons on gel filtration method. Optimal temperatures for both enzymes were $70^{\circ}C$ and optimal pH were about 6 and 10.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.243-247
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• 1994

Endo-polygalacturonase was purified from Jujube. The purification procedures included DEAE-cellulose ion exchange chromatography and gel filtration on Sephdex G-100. Enzyme was purified as a single protein band and purification yield was about 6%. When the purified enzyme was applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 19,000. Purified enzyme formed hexagonal board type.