• KSBB Journal
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• v.30 no.6
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• pp.291-295
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• 2015

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

• BMB Reports
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• v.34 no.3
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• pp.219-224
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• 2001

To identify the antioxidative peptides in the gelatin hydrolysate of bovine skin, the gelatin was hydrolyzed with serial digestions in the order of Alcalase, pronase E, and collagenase using a three-step recycling membrane reactor. The second enzymatic hydrolysate (hydrolyzed with pronase E) was composed of peptides ranging from 1.5 to 4.5 kDa, and showed the highest antioxidative activity, as determined by the thiobarbituric acid method. Three different peptides were purified from the second hydrolysate using consecutive chromatographic methods. This included gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an octadecylsilane chloride column. The isolated peptides were composed of 9 or 10 amino acid residues. They are: Gly-Glu-Hyp-Gly-Pro-Hyp-Gly-Ala-Hyp (PI), Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (PII), and Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp (PIII), as characterized by Edman degradation and fast-atom bombardment mass spectrometry. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability with a methylthiazol tetrazolium assay The results showed that PII had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by the addition of the peptide. These results suggest that the purified peptide, PII, from the gelatin hydrolysate of bovine skin is a natural antioxidant, which has potent antioxidative activity.

• Bulletin of the Korean Chemical Society
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• v.29 no.2
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• pp.342-346
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• 2008

Aurintricarboxylic acid (ATA) prevents apoptosis in a wide range of cell types, including PC12 cells. ATA is known to increase the phosphorylation level of IGF-1 receptor (IGF-1R) and downstream signaling proteins. ATA can translocate across the plasma membrane of PC12 cells and inhibit protein tyrosine phosphatases (PTPs) and, therefore, it is not clear whether ATA exerted its antiapoptotic effect through activation of IGF-1R or by inhibition of cytosolic PTPs. When PC12 cells, deprived of serum, were treated with Fab fragment of anti-IGF-1R antibody to prevent the binding of ATA to the extracellular domain of IGF-1R, ATA was found to penetrate into the cytosolic space of the cells. Under these conditions, the survival-promoting effects of ATA were abolished, and the increase of phosphorylation and characteristic cleavage of IGF-1R were not observed. These results indicate that the antiapoptotic effect of ATA in PC12 cells is due to the binding of ATA to the extracellular domain of IGF-1R and subsequent activation of the IGF-1R, not inhibition of cytosolic PTP(s).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.198-204
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• 2000

In order to utilize by-products which would normally be discarded in marine processing plants, cod teiset protein was hydrolyzed and antioxidative actiTity of the hydrolysate was investigated. AntioxidatiTe peptide was isolated using ultrafiltration membrane, ion-exchange chromatography on a SP-Sephadex C-25 column, gel filtration on a Sephadex G-15 column, high performance liquid chromatography on an ODS column, and capillary electrophoresis chromatography. Antioxidative activities of the cod teiset hydrolysate were compared with ${\alpha}-tocopherol$, one of the commercial antioxidant. The hydrolysate passed through a membrane with molecular weight cut-off (MWCO) 1 kDa was shown the strongest antioxidative activity, and the activity was higher $10{\%}$ as compared with ${\alpha}-tocopherol$. In addition, the peptide isolated by ion-exchange chromatography, gel filtration, and HPLC, respectively, was higher $53{\%}$ as compared with ${\alpha}-tocopherol$, and the amino acid sequence was Ser-Asn-Pro-Glu-Trp-Ser-Trp-Asn.

• Applied Chemistry for Engineering
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• v.3 no.4
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• pp.605-613
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• 1992

NCO-terminated prepolymers were synthesized by reacting carbonate-type polyol(PTMCG)($M_w=1,000$ and 2,000) with MDI and N-methyldiethanolamine, as a chain extender. Carbonate-type polyurethane containg zwitterionic group was prepared by reacting the prepolymer with 1,3-propane sultone. From the IR and NMR spectra of model reactions, it was known that the ionization occurred under the same condition. The structure of zwitterionic carbonate-type polyurethane(ZPU) therefore could be confirmed from the model reactions. Glass transition temperature(Tg) ranged between $-15{\sim}-30^{\circ}C$ from the thermal data. Tg was between $-15{\sim}-18^{\circ}C$ for a series of ZPU10 samples and between $-25{\sim}-26^{\circ}C$ for a series of ZPU20 polymers. Tensile strength increased with mole ratio of ionic content. On the contrary, elongation was rather dropped with mole ratio of ionic content. ZPU10-30 having better tensile strength and less elongation was selected as a membrane for the concentration of ethanol aqueous solution through pervaporation. To obtain the better selectivity, it was crosslinked with HMDI. In the swelling test, it showed the higher swelling degree at around 50wt% ethanol concentration due to the plastization effect of ethanol. To optimize the separation capacity, two operating factors-feed concentration and temperature-were considered. The overall separation capacity was as follows : separation factor, 2~83.2 ; the flux, $25.4{\sim}58.8g/m^2hr$.

• Applied Chemistry for Engineering
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• v.3 no.4
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• pp.595-604
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• 1992

Carbonate-type polyurethane resins containing anionic moieties were systhesized from NCO-terminated prepolymer method. Membranes were manufactured from the polymer solution and the separation of aqueous ethanol solution was investigated. To enhance the property of urethane resin, carbonate-type polyol(PTMCG) was used. ${\alpha}^{\prime},{\alpha}^{{\prime}{\prime}}$-dimethylolpropionic acid was used as a chain extender to increase the hydrophilicily of the urethane membrane. The ionization of the pendent carboxylic groups in urethane resin was carried out using trimthylamine. To confirm the formation of anionic groups in urethane resin, IR spectra of model compounds were compared with those of urethane resins. It was confirmed that the concentration of hard segment and hydrogen bond contributed to the property of the concentration of hard segment and hydrogen bond contributed to the property of urethane resin in which the mole ratio of chain extender and polyol was from 3:1 to urethane resin in which the mole ratio of chain extender and polyol was from 3:1 to 5:1. The carbonate-type polyurethane containing pendent carboxylic grop(PU) had Tg of around-$25^{\circ}C$ and Tm, $45^{\circ}C$ measured by DSC. Transition temperatures of one containing pendent anionic group(APU) prepared from the ionization of PU shifted to $8{\sim}10^{\circ}C$ lower temperature region than those of PU. Pervaporation membrane was prepared through the casting method. N, N-dimethylformamide (DMF) were used as a solvent and hexamethylene diisocyanate(HMDl) as a crosslinking agent. Swelling degree increased with ethanol concentration in mixure and the control of the swelling degree of the membrane could be achieved by crossliking. The results of pervaporation were as follows : separation factor, 2.3~9.8 ; flux, $27{\sim}79.5g/m^2hr$. Pervaporation separation capacity could be enhanced by reducing the molecular weight of polyol from 2,000 to 1,000.

• Applied Chemistry for Engineering
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• v.19 no.5
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• pp.477-483
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• 2008

$La_{0.7}Sr_{0.3}Co_{0.2}Fe_{0.8}O_{3-{\delta}$ oxide was synthesized by a citrate method and a typical dense membrane of perovskite oxide has been prepared using as-prepared powder by pressing and sintering at $1300^{\circ}C$. Precursor of $La_{0.7}Sr_{0.3}Co_{0.2}Fe_{0.8}O_{3-{\delta}$ prepared by citrate method was investigated by TGA and XRD. Metal-citrate complex in precursor was decomposed into perovskite oxide in the temperature range of $260{\sim}410^{\circ}C$ but XRD results showed $SrCO_3$ existed as impurity at less than $900^{\circ}C$. Electrical conductivity of membrane increased with increasing temperature but then decreased over $700^{\circ}C$ in air atmosphere ($Po_2=0.2atm$) and $600^{\circ}C$ in He atmosphere ($Po_2=0.01atm$) respectively due to oxygen loss from the crystal lattice. The oxygen permeation flux increased with increasing temperature and maximum oxygen permeation flux of $La_{0.7}Sr_{0.3}Co_{0.2}Fe_{0.8}O_{3-{\delta}$ membrane with 1.6 mm thickness was about $0.31cm^3/cm^2{\cdot}min$ at $950^{\circ}C$. The activation energy for oxygen permeation was 88.4 kJ/mol in the temperature range of $750{\sim}950^{\circ}C$. Perovskite structure of membrane was not changed after permeation test of 40 h and the membrane was stable without secondary phase change with 0.3 mol Sr addition.

• Membrane Journal
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• v.29 no.6
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• pp.348-354
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• 2019

Recently, the application range of organic solvent nanofiltration (OSN) technology has been expanding, requiring membranes with better performance. In this work, thin film composite (TFC) OSN membrane was fabricated. First, ultrafiltration support membrane was prepared via nonsolvent-induced phase separation (NIPS) technique using polysulfone (PSf) and polyethersulfone (PES). Then, the effect of pore forming additives such as polyvinylpyrrolidone (PVP) and pluronic F-127 were employed to improve the membrane permeance. The well-known interfacial polymerization technique was employed using MPD-TMC chemistry to form a thin film on top of the fabricated support, and its solvent permeance and nanofiltration performance was characterized. It was found that polyethersulfone support exhibited more reliable performance compared to polysulfone, and PVP additive was more effective compared to Pluronic F-127. As for the oSN performance, polar aprotic solvents like acetonitrile show significantly higher flux (986.5 L·m^-2·h^-1·bar^-1) compared to water and EtOH (9.5 L·m^-2·h^-1·bar^-1).

• Bulletin of the Korean Chemical Society
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• v.32 no.5
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• pp.1645-1649
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• 2011

[ ${\beta}$ ]Ketoacyl acyl carrier protein synthase I (KAS I) is involved in the elongation of unsaturated fatty acids in bacterial fatty acid synthesis and a therapeutic target of designing novel antibiotics. In this study, we performed receptor-oriented pharmacophore-based in silico screening of E. coli KAS I (ecKAS I) with the aim of identifying novel inhibitors. We determined one pharmacophore map and selected 8 compounds as candidates ecKAS I inhibitors. We discovered one antimicrobial compound, YKAe1008, N-(3-pyridinyl) hexanamide, displaying minimal inhibitory concentration (MIC) values in the range of 128-256 ${\mu}g/mL$ against MRSA and VREF. YKAe1008 was subsequently assessed for binding to ecKAS I using saturation-transfer difference NMR spectroscopy. Further optimization of this compound will be carried out to improve its antimicrobial activity and membrane permeability against bacterial cell membrane.

• Journal of Korean Society of Water and Wastewater
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• v.21 no.3
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• pp.349-357
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• 2007

Reports of endocrine disrupting compounds (EDCs), pharmaceutically active compounds (PhACs), and personal care products (PCPs) have raised substantial concern in important potable drinking water quality issues. Our study investigates the removal of EDCs, PhACs, and PCPs of 10 compounds having different physico-chemical properties (e.g., molecular weight, and octanol-water partition coefficient ($K_{OW}$)) by nanofiltration (NF) membranes. The rejection of micropollutants by NF membranes ranged from 93.9% to 99.9% depending on solute characteristics. A batch adsorption experiments indicated that adsorption is an important mechanism for transport/removal of relatively hydrophobic compounds, and is related to the octanol-water partition coefficient values. The transport phenomenon associated with adsorption may also depend on solution water chemistry such as pH and ionic strength influencing the pKa value of compounds. In addition, it was visually seen that the retention was somewhat higher for the larger compounds based on their molecular weight. These results suggest that the NF membrane retains many organic compounds due to both hydrophobic adsorption and size exclusion mechanisms.