• Journal of Microbiology
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• 제35권2호
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• pp.134-140
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• 1997

Botrytis cinerea T91-1 has been shown to produce at least four different polygalacturonases into a liquid medium containing citrus pectin, a carbon sousrce. One of the enzymes, which had an apparent molecular weight of 66 kDa estimated by denatured polyacrylamide gel electrophoresis, was purified to electrophoretic homogeneity by a series of procedures including a cetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. The molecular weight of native enzyme was determined to be 64 kDa by gel permeation chromatography indicating the enzyme to be a single polypeptide chain. By viscometric analysis, the enzyme was revealed as exo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Mg^{2+}$, and Cu$^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was 5$0^{\circ}C$. And the enzyme showed optimal pH values between 4.0 and 5.0. The enzyme was stable upto 12 hours in the range of pH 3 to 8 and at temperature below 3$0^{\circ}C$.

• 한국식품영양학회지
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• 제24권3호
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• pp.291-294
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• 2011

A lipase inhibitor from Desmodium oxyphyllum DC. was purified by methanol extraction, systematic solvent extraction, silica gel column chromatography, $C_{18}$ solid phase extraction chromatography and RP-HPLC. We obtained the purified lipase inhibitor with 182 ng($IC_{50}$) of lipase inhibitory activity for a 0.06% yield. Its molecular weight was estimated to be 655.37 Da from an instrumental analysis of MALDI-TOF-MS and it was identified copper-3,5-dibromo-2-hydroxybenzoic acid ($C_{14}H_8Br_4CuO_6$) by $^1H$, $^{13}C$ NMR analysis.

• 한국표면공학회지
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• 제40권1호
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• pp.16-22
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• 2007

Recently, mesoporous metallic materials are becoming more and more important in various applications like catalysts, electrochemical detectors, batteries, and fuel cells because of their high surface area. Among the various methods for manufacturing mesoporous structure, surfactant templating method followed by electroplating has been tried in this study. A mesoporous metallic film was prepared by electrodeposition from electroplating solution mixed with surfactant template. Nonionic type lyotropic liquid crystalline surfactant, Brij56, and nickel acetate based solution were selected as a template material and electroplating solution, respectively. To determine the content of surfactant forming a hexagonal column structure, the phase diagram of electroplating solution and surfactant mixture has been exploited by polarized optical microscopy equipped with heating and cooling stage. Nickel films were electroplated on Cu foil by stepwise potential input method to alleviate the concentration polarization occurred during the electroplating process. TEM and XRD analyses were performed to characterize the size and shape of mesostructures in manufactured nickel films, and electrochemical characterization was also carried out using cyclic voltammetry.

• Ocean and Polar Research
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• 제28권3호
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• pp.215-224
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• 2006

Spatial variation of heavy metal accumulation was investigated in Manila clam Ruditapes philippinarum collected from several tidal flats. Sediment metal levels varied highly among the sites, which was attributed primarily to differences in Fe and organic carbon contents and in part to gain size. Significant differences in metal concentrations also were found in the clam tissue among the different sampling sites. However, except for a few metals (Mn, Zn, Pb), which showed some elevation, the variations in the clam tissue were not related to the variations in the sediment. This is likely because most metals in filter-feeding herbivores such as R. philippinarum accumulated as a result of feeding on suspended particles such as phytoplankton and organic detritus in the water column, not in bottom sediment. In addition, tissue weight for a specific shell size varied significantly among the sites, and increased tissue mass indicating a good nutritive condition likely caused a subsequent dilution of body metals leading to reduced weight-specific concentrations of some metals (Cd, Zn, Cu, Co).

• 한국미생물·생명공학회지
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• 제24권2호
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• pp.197-205
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• 1996

The $\beta$-xylosidase was purified 99- fold from the culture supernatant of Pseudo onas sp. CB-33 by ammonium sulfate precipitation, PEI precipita- tion, DEAE-Sephadex column chromatography, Sephadex G-75 gel filtration chromatography and preparative disc gel electrophoresis. Molecular weight of the enzyme was estimated to be 44,000 by SDS polyacrylamide gel electrophoresis. The enzyme has a pH optimum for activity at 7.0 and is stable over pH 6.5-9.0. The optimal temperature of the enzyme was 45$\circ$C, and its enzymatic activity was completely inactivated at 55$\circ$C for 30 min. Km value of the enzyme for p-nitrophenyl-$\beta$-D-xylopyranoside was calculated to be 4.6 mM. The effect of various reagents on the $\beta$-xylosidase activity was investigated. The enzyme activity was completely inhibited by Hg$^{2+}$, Cu$^{2+}$ and Zn$^{2+}$. The $\beta$-xylosidase was inactivated by tryptophan-specific reagent, N-bromosuccinimide and tyrosine-specific reagent, iodine. The enzyme could degrade xylo-oligosaccharides to xylose and the enzyme was competitively inhibited by xylose. The $\beta$-xylosidase and endoxylanase from Psedomonas sp. CB-33 hydrolized xylan synergically. The purified enzyme also showed $\alpha$-L-arabinofuranosidase activity.

• 미생물학회지
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• 제14권2호
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• pp.84-94
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• 1976

Cellulase components, CMCase(Cx) and Avicelase$(C_1)$, were partially prueified, from the culture extract of a strain of Trichoderma koningii by column chromatography on DEAE-Sephadex A-50 and the step of gel filtration through Sphadex G-150, Optimum pH of CMCase was 5.2 and Avicelase showed the highest activity at pH 5.6 in acetate buffer. Optimal temperatures for activities of CMCase and Avicelase were $50^{\circ}C\;and\;60^{\circ}C, $ respectively. More than 70% of the activities of two enzymes were remained after heating at $60^{\circ}C$ for 30 min and Avicelase is more stable than any other fungal enzymes. The Michaelis constants, Km, of CMCase and Avicelase were 0.116% of CMC and 0.281% of avicel. And also the values of maximum velocity, Vmax, of CMCase and Avicelase were $23.20{\mu}g\;and\;2.54{\mu}g$ of reducing sugar per min. Of the metal ions tested against the activites of CMCase and Avicelase, $Cu^{++}, \; Hg^{++}, \;and\;Pb^{{++}$ are remarkably effective inhibitors. The molecular weights of Cx and $C_1$ component were estimated to be about 47, 000 and 61, 000 by gel filtration method.

• Fisheries and Aquatic Sciences
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• 제13권1호
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• pp.49-55
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• 2010

A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

• 한국식품영양과학회지
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• 제24권4호
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• pp.563-569
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• 1995

Myrosinase from Korean cabbage(Bogdoli) was purified and its enzymatic properties were investigated. Myrosinase from the Korean cabbage was purified by DEAE Bio-Gel Sepharose, Concanavalin-A, and Mono-Q column chromatography and exhibited a 55KD molecular weight with a single band on the gel of SDS-PAGE. The enzyme was purified about 21-fold compared to its crude enzyme and a specific activity of purified enzyme was 15, 120units/mg. Optimum pH of the myrosinase was 7.0 in both phosphate and Tris-HCl buffer solutions, the enzyme was stable at pH 6.5~7.0. Optimum temperature of enzyme was 37~38$^{\circ}C$. The enzyme activity was significantly inhibited by Cu2+ and Hg2+, but enhanced by ascorbic acid, resulting in a maximum activity at 1mM ascorbic acid. Among the ascorbic acid analogues, dehydro-ascorbic acid did not affect, whereas others showed a little effect on the enzyme activity, but less than ascorbic acid itself. Reducing agents such as 2-mercaptoethanol and dithiothreitol had no effect on the enzyme activity, but the enzyme activity was enhanced when 2-mercaptoethanol was mixed with ascorbic acid.

• Advances in environmental research
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• 제3권4호
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• pp.283-292
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• 2014

Iron precipitating organisms play a significant role in the formation of ferric hydroxide precipitate, which acts as strong adsorbent for toxic metal. In this respect four different iron precipitating cultures were isolated from Hutti gold mine surface winze water sample on citrate agar medium. The best isolate was screened out for metal removal study on the basis of fast visual iron precipitation. The selected isolate was identified as Enterobacter sp. based on routine biochemical tests and Biolog GN microplate results and as Enterobacter cloacae subsp. dissolvens by 16S rRNA gene sequence analysis (GenBank accession number EU429448). Influence of medium composition, medium initial pH, the influence of inoculum size, effect of various media and ferric ammonium citrate concentration were studied on metal removal in shake flask experiments. Under the optimized conditions studied, E. cloacae showed $94{\pm}2$, $95{\pm}2$ and $70{\pm}2%$ of cadmium, copper and mercury removal from a simulated waste in shake flask studies. In lab scale column reactor more than 85% of copper and mercury removal was achieved.

• 한국정밀공학회지
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• 제21권11호
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• pp.186-194
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• 2004

In this research, microfabrication technique using localized electrochemical deposition (LECD) with ultra short pulses is presented. Electric field is localized near the tool tip end region by applying a few hundreds of nano second pulses. Pt-Ir tip is used as a counter electrode and copper is deposited on the copper substrate in 0.5 M CuSO$_4$ and 0.5 M H$_2$SO$_4$ electrolyte. The effectiveness of this technique is verified by comparison with LECD using DC voltage. The deposition characteristics such as size, shape, surface, and structural density according to applied voltage and pulse duration are investigated. The proper condition is selected from the results of the experiments. Micro columns less than 10 $\mu$m in diameter are fabricated using this technique. The real 3D micro structures such as micro pattern and micro spring can be fabricated by this method. It is suggested that presented method can be used as an easy and inexpensive method for fabrication of microstructure with complex shape.