• Journal of Life Science
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• v.11 no.6
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• pp.561-567
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• 2001

To isolate and purify fibrinolytic active substance from Sarcodon aspratus(N $H_4$)$_2$S $O_4$ precipitation, DE52 anion exchange column chromatography, Sephacryl-S 200gel filtration chromatography and Mono S cation FPLC were carried out and the characterizations of the purified enzyme were investigated. The bound active fraction on DE52 anion exchange column chromatography were eluted with 0.2 M NaCI and the fibrionlytic enzyme was purified after following Sephacryl-S200 gel fitration chromatography and Mono S cation EPLC. The specific activity of purified enzyme was 55.2 U/mg protein and increased 11.3 fold comparing crude extract and the yield was 49.5%. 12% SDS-PAGE electrophoresis and gel filtration chromatography revealed that Sarcodon aspratus fibrionloytic enzyme was highly purified and had 29.300 Da molecular weight. Enzyme activity of the purified fibrinolytic enzyme from Sarcodon aspratus was increased on higher pH and was stable until pH 10.5. On temperature dependent stability, the enzyme activity was decrease sharply but remained 25% relative activity on 8$0^{\circ}C$. This enzyme activity was inhibited by heavy metal ion, C $U^{2+}$ and $Co^{3+}$ with 68% and 38%, respectively. And also, the enzyme activity was inhibited with $Ca^{2+}$ chelator EDTA and serine protease inhibitor PMSF. These results from this study suggested that the fibrinolycit enzyme from Sarcodon aspratus is a serine protease and the enzyme activity was increased by $Ca^{2+}$ or $Mg^{2+}$ ion.n.ion.n.

• Bulletin of the Korean Chemical Society
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• v.26 no.4
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• pp.569-574
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• 2005

A coupled column liquid chromatography system was applied for the separation of the burnup monitors in spent nuclear fuel sample solutions. A reversed phase column was studied for the adsorption behavior of uranyl ions using alpha-hydroxyisobutyric acid as an eluent and used for the separation of plutonium and uranium. A cation exchange column prepared by coating 1-eicosylsulfate onto the reversed phase column was used for the separation of the lanthanides. In addition, retention of Np was checked with the reversed phase column and cation exchange column, respectively, according to the oxidation states to observe the interference effect for the separation of burnup monitors. This chromatography system showed a great reduction in separation time compared to a conventional anion exchange method. A good agreement from the burnup data was obtained between for this method and a conventional anion exchange method to within 1% of a difference for the spent nuclear fuel samples of about 40 GWD/MTU.

• Analytical Science and Technology
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• v.13 no.4
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• pp.446-452
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• 2000

The simulated nuclear spent fuel (SIMFUEL) containing the platinum group elements which will not be dissolved in a nitric acid was completely dissolved with a acid digestion bomb. The metallic elements separated in the SIMFUEL were measured by inductively coupled plasma atomic emission spectrometry (ICP-AES). Because the peaks of metallic elements were spectrally interfered by uranium spectrum, uranium and metallic elements were separated by cation exchange resin for Mo, Pd, Rh and Ru and by anion exchange resin for Ba, Ce, La, Nd, Rh, Sr, Y and Zr, respectively. The recovery of Mo, Pd, Rh and Ru after separation by cation exchange chromatography found to be 99-103% and anion exchange separation showed 96.5-107% of recovery except Y with the simulated solution whose concentration was similar to the spent nuclear fuel. The relative standard deviation of this method showed 1.3-6.7% in the SIMFUEL whose concentrations of metallic elements were between several $10^2-10^3$ppm.

• Bulletin of the Korean Chemical Society
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• v.33 no.10
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• pp.3298-3302
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• 2012

Although offline enrichment of phosphorylated peptides is widely used, enrichment for phosphopeptides using $TiO_2$ is often performed manually, which is labor-intensive and can lead to irreproducible results. To address the problems associated with offline enrichment and to improve the effectiveness of phosphopeptide detection, we developed an automated online enrichment system for phosphopeptide analysis. A standard protein mixture comprising BSA, fetuin, crystalline, ${\alpha}$-casein and ${\beta}$-casein, and ovalbumin was assessed using our new system. Our multidimensional system has four main parts: a sample pump, a 20-mm $TiO_2$-based column, a weak anion-exchange, and a strong cation-exchange (2:1 WAX:SCX) separation column with LC/MS. Phosphorylated peptides were successfully detected using the $TiO_2$-based online system with little interference from nonphosphorylated peptides. Our results confirmed that our online enrichment system is a simple and efficient method for detecting phosphorylated peptides.

• Journal of the Korean Chemical Society
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• v.41 no.11
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• pp.612-638
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• 1997

The methods of separation and recovery of rare earth elements in monazite sand have been studied by the ion exchange chromatography. Both of cation and anion exchange resin were used as ion exchange resins and the solutions of EDTA, DTPA, IMDA and Ln-EDTA were used as eluents. The H+, Zn2+, Fe3+, Al3+, Cu2+, and NH4+ forms of cation exchange resin were used as retaining ions. Ln-EDTA solution was loaded on the EDTA form of anion exchange resin and separated. The Ln-EDTA solution was also used as an eluent for a selective separation of one element from the rare earth mixture solution. The size effects of resin column, the elution mechanism for the various elution types and the separation of a large amount of rare earths were studied.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.861-869
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• 1999

An extracellular protease was purified to homogeneity from the culture supernatant of psychrotrophic bacteria Bacillus sp. JH 108 using procedures including ammonium sulfate fractionation, anion exchange chromatography, gel filtration chromatography, and cation exchange chromatography. The enzyme exhibited a molecular weight of 36 kDa, an optimum pH of 8 to 9, and optimum temperature of $60^{\circ}C$. The enzyme preferentially hydrolyzed leucine at the N-terminus of peptides and thus can be classified as an aminopeptidase. It was strongly inhibited by metal chelating agents such as EDTA and l, l0-phenanthroline. The activity lost by EDTA was restored with $Zn^{2+}{\;}or{\;}Co^{2+}$. These divalent cations also stimulated the native enzyme. This suggests that the enzyme is a metalloprotease acting as a leucine aminopeptidase.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.117-123
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• 1989

Radioactive N-$\alpha$-p-nitrobenzoxycarbonyl (NBZ)-L-[2,$3-^{3}$H] arginyl diazomethane was used as an affinity label for the vacuolar arginine transporter in Neurospora crassa. Vacuolar matrix proteins were removed by fracturing the membranes with freeze-thaw method in dry ice/ethanol bath. Vacuolar membrane proteins were then wasged with 500mM NaCl to remove ionically bound derivatives and peripheral membrane proteins from vacuolar membranes. After dissolved in 1% Titon X-100, dissolved vacuolar memvrane proteins were separated with molecular sieve column chromatography, anion and cation exchange chromatographies. The arginine transporter was purified giving the purification factor of 1136.

• Environmental Analysis Health and Toxicology
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• v.1 no.1
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• pp.77-80
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• 1986

Radiochemical synthesis of $N^{G}$-mono［$^{14}$ C-methyl］-L-arginine is described. The compound was synthesized from radio-active mono［$^{14}$ C］-methylamine as easily and purified by strong cation-exchange resin (NH form) liquid chromatography using a gradient of ammonium hydroxide, and crystallized as flavianate. The free amino acid was successfully prepared by strirring its flavianate and strong anion-exchange resin (OH- form), which could remove the flavianic acid from its salt in water below room temperature. Purity of the compound was tested by thin-layer chromatography, thin-layer electro-phoresis, and scintillation spectrometry.y.

• KSBB Journal
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• v.15 no.6
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• pp.605-608
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• 2000

Oligodeoxynucleotides (ODNs) were separated by high-performance membrane chromatography (HPMC), a combined system of chromatography and membrane. The separation mechanism involved anion-exchange, and the stationary phase was cation CIM (Convective Interaction Media) DEAE disk (16${\times}$3 mm). Two types of mobile phase were used, buffer A (20mM Tris-HCl, pH 7.4) and buffer B (buffer A + 1M NaCl). As the amount of NaCl dissolved in buffer linearly increased, the retention time shortened, which enabled a gradient elution mode. Based on the number of theoretical plates and resolution observed, the optimum mobile phase and operating condition (Buffer A/Buffer B=50/50 - 20/80 vol%, gradient time 2 min) were experimentally determined. In this experimental condition, ODNs were separated within 2 min at a mobile phase flow rate of 6 ml/min.

• KSBB Journal
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• v.16 no.6
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• pp.533-537
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• 2001

${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin in whey proteins were separated by high performance membrane chromatography (HPMC). The separation mechanism involved anion-exchange, and the stationary phase was anion CIM (Convective Interaction Media) DEAE, QA disk and cation exchanger SO$_3$(16${\times}$3 mm). Two types of mobile phase were used, buffer A (20 mM Tris-HCI, pH 7.3) and buffer B(buffer A + 1 M NaCl), As the amount of NaCl dissolved in buffer linearly increased, which enabled a gradient elution mode. The optimum mobile phase and operating condition (Buffer A/Buffer B = 100/0 - 30/70 vol%, gradient time 1 min, 30/70 - 10/90 vol.%, gradient time 2 min) were experimentally determined. In this experimental condition, ${\alpha}$-lacta1bumin, ${\beta}$-lactoglobulin were separated within 5 min at a mobile phase flow rate of 4 mL/min.