• Molecules and Cells
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• v.21 no.1
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• pp.29-33
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• 2006

We have established three immortal bovine muscular epithelial (BME) cell lines, one spontaneously immortalized (BMES), the second SV40LT-mediated (BMEV) and the third hTERT-mediated (BMET). The morphology of the three immortal cell lines was similar to that of early passage primary BME cells. Each of the immortal cell lines made cytokeratin, a typical epithelial marker. BMET grew faster than the other immortal lines and the BME cells, in 10% FBS-DMEM medium, whereas neither the primary cells nor the three immortal cell lines grew in 0.5% FBS-DMEM. The primary BME cells and the immortal cell lines, with the exception of BMES, made increasing amounts of p53 protein when treated with doxorubicin, a DNA damaging agent. On the other hand, almost half of the cells in populations of the three immortal cell lines may lack $p16^{INK4a}$ regulatory function, compared to primary BME cells that were growth arrested by enforced expression of $p16^{INK4a}$. In soft-agar assays, the primary cells and immortal cell lines proved to be less transformed in phenotype than HeLa cells. The three immortal epithelial-type cell lines reported here are the first cell lines established from muscle tissue of bovine or other species.

• Journal of Life Science
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• v.12 no.1
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• pp.77-86
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• 2002

Chitin, a $\beta$-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl rhitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnS $O_4$.4$H_2O$, 37$^{\circ}C$, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pI) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and $65^{\circ}C$, respectively. Chi-56A was stable up to $65^{\circ}C$ and in alkaline region. Its $K_{m}$ value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.e.

• Korean Journal of Food Science and Technology
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• v.40 no.2
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• pp.220-227
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• 2008

In this study, polysaccharides were isolated from Korean persimmon vinegar to characterize the polysaccharides existing as soluble forms within traditional Korean fermented beverages, and their immuno-stimulating activities were examined. Three successive chromatographies were used to purify the main polysaccharide in the persimmon vinegar, PV-1b-I, to homogeneity from the crude polysaccharide (PV-0). The molecular mass of PV-1b-I was estimated as 110 kDa and it contained significant proportions of mannose (46.8%), galactose (28.5%) and arabinose (19.1%). PV-1b-I strongly reacted with ${\beta}$-glucosyl Yariv reagent, suggesting the presence of an arabino-3,6-galactan moiety. PV-1b-I also induced high levels of macrophage activation and mitogenicity on murine splenocytes in vitro. The intravenous administration of PV-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. PV-1b-I also showed potent anticomplementary activity in a dose-dependent manner. Finally, C3 activation products were identified by crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of PV-1b-I under $Ca^{2+}$-free conditions, suggesting that this PV-1b-I causes complementary activations via both alternative and classical pathways. From these results, one can conclude that Korean persimmon vinegar contains select polysaccharides in addition to healthy components, and these polysaccharides appear to provide immuno-stimulating activities beneficial to human health.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.552-558
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• 2002

Polyphenol oxidase from Flammulina velutipes was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Superdex G-200 gel filtration chromatography, Phenyl superose affinity chromatography, Mono-Q anion exchange chromatography and Superdex S-200 gel filtration chromatography on FPLC. After these purification steps specific activity of purified polyphenol oxidase increased to 199.1 units/mg. Polyphenol oxidase from F. velutipes was composed of a single polypeptide with molecular weight of about 40 kDa. Optimum pH and temperature for the enzyme reaction were found to be 6.0 and $25^{\circ}C$, respectively. The activity of the enzyme gradually decreased at acidic pH between 3 and 5, and the enzyme lost its activity at alkaline pH between 8 and 10. This enzyme exhibited high substrate specificity to o-diphenols. Km-values for L-DOPA and caffeic acid were found to be 3.97 mM and 1.78 mM, respectively. 2-mercaptoethanol, L-ascorbic acid, sodium bisulfite, EDTA and $Mg^{2+}$ inhibited the activity of pholyphenol oxidase and $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$ and $Ni^{2+}$ increased enzyme activity. The activity of enzyme was well maintained at $-70^{\circ}C$ for over 4 months, and at $-20^{\circ}C$ for 1 months.

• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1158-1167
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• 2000

To produce ${\beta}-cyclodextrin({\beta}-CD)$, a cyclodextrin glucanotransferase(CGTase) producing Aspergillus sp. CC-2-1 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of CGTase reached to the maximum when the wheat bran medium containing 0.1% albumin, 2% $(NH_4)_2S_2O_8$, 2% soluble starch and 0.2% $KH_2PO_4$ was cultured for 5 days at $37^{\circ}C$. The purity of CGTase was increased by 13.14 folds after DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and the specific activity was 172.14 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of CGTase was estimated to be 27,800 by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the CGTase activity were 9.0 and $80^{\circ}C$, respectively. The enzyme was stable in pH $8.0{\sim}11.0$ at $60{\sim}80^{\circ}C$. The activity of purified enzyme was activated by $K^+,\;Cu^{2+}$ and $Zn^{2+}$. The activity of the CGTase was inhibited by the treatment with 2,4-dinitrophenol and iodine. The result suggests that the purified enzyme has phenolic hydroxyl group of tyrosine, histidine imidazole group and terminal amino group at active site. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the $K_m$ value of 18.182 g/L with the $V_{max}$ of 188.68 ${\mu}mole/min$. The activation energy for the CGTase was calculated by Arrhenius equation was 1.548 kcal/mol.

• Korean journal of food and cookery science
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• v.19 no.5
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• pp.616-623
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• 2003

Waxy barley brans were collected during the pearling process. The extraction of $\beta$-glucan from barley bran was effected by the extraction conditions. The $\beta$-glucan content increased with temperature, but not with pH. The highest yield, 6.5%, was achieved at pH 7.0 and 55$^{\circ}C$. At pH 10 and 45$^{\circ}C$, 48.5% of the $\beta$-glucan in barley bran was recovered in the gum product, with 54.6% purity. The protein and starch contaminations were high, reaching 13.6 and 23.7%, respectively. The $\beta$-glucan content was greatest in the subaleurone and aleurone regions (bran fractions 1, 2, 3 and 4), and declined considerably toward the inner layers. A monosaccharide analysis of the purified, $\beta$-glucan, from bran fractions 1, 2, 3 and 4, indicated that glucose constituted the majority of the gum. The small amounts of the arabinose and xylose found in the gum may indicate the presence of arabinoxylans as minor constituents. The molecular weights of the $\beta$-glucans isolated from bran fractions 1,2 and 3 were found to be 4.09${\times}$10$^{5}$ ∼-4.41${\times}$10$^{5}$ . The major glycosidic linkages of the $\beta$-glucans demonstrated the presence of 2, 4, 6-Me-Glc and 2, 3, 6-Me-Glc. When flow behaviors of barley bran $\beta$-glucan were examined, $\beta$-glucan exhibited pseudoplastic fluid properties.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.5
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• pp.769-777
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• 2004

A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70％ acetone extract, showing high stability against heat treatment and peptic digestion.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.171-179
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• 2013

Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

• Polymer(Korea)
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• v.38 no.1
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• pp.43-48
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• 2014

In this paper, we made a new polycationic polymeric liposome composed of low molecular weight polyethylenimine (PEI) and 10,12-pentacosadiynoic acid (PCDA). PCDA liposome was prepared by ultraviolet irradiation. PEI was further conjugated on the surface of the polymerized PCDA liposome using coupling reagents to make PCDA-PEI. The blue-to-red transition of PCDA liposome was observed during the coupling reaction. The size distribution of liposome and complexes with plasmid DNA was measured by dynamic light scattering (DLS). The complex formation was also identified by agarose gel electrophoresis and PicoGreen reagent assay. We confirmed the complex formation of the polymeric liposome with DNA and then performed transfection and cytotoxicity assay in HEK 293 and HeLa cells. As a result, PCDA-PEI showed significant gene transfection efficiency and low cytotoxicity. This study shows that PEI-conjugated PCDA liposome could be an efficient gene or drug delivery carrier.

• Polymer(Korea)
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• v.38 no.1
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• pp.31-37
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• 2014

To prepare biomass based nylon 6,5 copolymers, ${\varepsilon}$-caprolactam and 2-piperidone, the monomers of nylon 6,5 copolymers, were synthesized respectively from lysine and 5-aminovaleric acid which were produced from glucose by the fermentation process. The copolymers were then polymerized by the anionic ring opening polymerization of them at $40^{\circ}C$, using potassium tert-butoxide as a catalyst and acetyl-2-caprolactam and carbon dioxide as initiators. The prepared copolymers were characterized with various analytical methods: their viscosity molecular weight ($M_{\eta}$) was as high as 30000 g/mol and polymerization yield was over 50%, and it was found that they were semi-crystalline polymers having melting point at $165^{\circ}C$ which was much lower than its thermal degradation point, $250^{\circ}C$. These polymers were expected to have good thermal processability and application fields.