• YAKHAK HOEJI
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• v.46 no.1
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• pp.24-29
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• 2002

Enoyl-Acyl Carrier Protein Reductase (Fabl) of bacteria is hem as an important target for new antibacterial drugs and plays a determinant role in completing cycles of elongation in type-H fatty acid synthase system. In this study, a fabI gene from Staphylococcus aureus 6538p cloned in pET-l4b vector and FabI protein was over-produced in Escherichaia coli BL2l (DE3). $NH_2$-terminal His-tagged FabI protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metalaffinity chromatography Purified 6xHis-tagged FabI showed a catalytic activity on tram - 2 - octenoyl - N -acethlcysteamine by utilizing NADPH as a cofactor. For the discovery of new FabI inhibitors from chemical libraries, a target-oriented screening system using a 96-well plate was developed. About 10,000 chemical libraries from Korea Chemical Bank wore tested in this screening system, and 26 chemicals (0.25%) among them showed an inhibitory activity against FabI enzyme. This result showed that a new screening system can be used for the discovery of new FabI inhibitors.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.978-983
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• 2004

The effects of glycyrrhizic acid (GLZ) on protein binding of diltiazem, verapamil, and nifedipine were investigated. Protein binding studies (human serum, human serum albumin (HSA) and (X1-acid glycoprotein (AAG)) were conducted using the equilibrium dialysis method with and without addition of GLZ. The binding parameters, such as the number of moles of bound drug per mole of protein, the number of binding sites per protein molecule, and the association con-stant, were estimated using the Scatchard plot. The serum binding of nifedipine, verapamil, and diltiazem was displaced with addition of GLZ, and the decreases of Ks for serum were observed. GLZ decreased the association constants of three drugs for HSA and AAG, while the binding capacity remained similar with addition of GLZ. Although the characteristics of interaction were not clear, GLZ seemed to mainly affect HSA binding of nifedipine rather than AAG binding, while GLZ seemed to affect both AAG- and HSA-bindings of verapamil and dilt-iazem resulting in a serum binding displacement.

• Applied Biological Chemistry
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• v.36 no.3
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• pp.191-196
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• 1993

This method using the synthesis substrate of $5-bromo-4-chloro-3-indolyl-{\alpha}-galactoside\;(X-{\alpha}-Gal)$ was examined for the differential enumeration of Bifidobacteria and lactic acid-producing bacteria. Bifidobacteria possess a high level of ${\alpha}-galactosidase$ activity. Bifidobacterium longum KCTC 3215 exhibited the highest ${\alpha}-galactosidase$ specific activity (8.57 units/mg protein). Determination of ${\alpha}-galactosidase$ activity using the PNPG procedure showed that Lactobacillus, Streptococcus, Pediococcus, and Leuconostoc strain had lower ${\alpha}-galactosidase$ activity as compared to Bifidobacteria. The $X-{\alpha}-Gal$ based medium is useful to identify Bifidobacteria among lactic acid-producing bacteria since the enzyme action of ${\alpha}-galactosidase$ spills $X-{\alpha}-Gal$ substrate and releases indol which impacts a blue color to Bifidobacterial colonies on agar plates. All strains of Bifidobacteria appeared as blue colonies on MRS agar medium supplemented with $100\;{\mu}M\;X-{\alpha}-Gal$ while colonies of other lactic acid-producing bacteria appeared white or light blue.

• The Korean Journal of Food And Nutrition
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• v.30 no.5
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• pp.1068-1079
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• 2017

This study was applied to the PCA (Primary Components Analysis) for the sixteen table setting at the 2017 Yeongju local food contest. In this contest, we have developed a seonbibansang and a temple one-dish meal. As a result of the correlation analysis, the applicability and composition were 0.7980, harmony and taste were 0.7747 and easiness and composition were 0.7435. In the Primary Component $Y_1$, all the variables $X_1{\cdots}X_{10}$ mean that the quality of the food had positive values greater than zero. The second Primary Component $Y_2$ has a large positive value while $X_4$, $X_5$, $X_6$, $X_7$, $X_9$ have negative values. $Y_2$ is a value representing the sanitation variable, and can be considered a traditional and characteristic table setting natural to the native food in Yeongju. In addition, we developed an-hyangbansang and seonmyoaecheong food content by applying PCA factors (the elements of harmony, ease and sanitation). Table setting of an-hyangbansang provided energy 61.5%, protein 20.0% and fat 18.5% and seonmyoaecheong provided energy 62.7%, protein 15.4% and fat 22.2%. This satisfied the necessary amount of caloric nutrient intake that could be provided in a meal. Especially through story-telling, a modern interpretation - or rebranding - of local and traditional foods could make these traditional food products familiar to consumers currently. The developed table setting is felt to be conductive to the possible commercialization and introduction of traditional food into the mainstream commercial food service industry.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1542-1546
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• 2009

Zeaxanthin glucosyltransferase (CrtX) mediates the formation of zeaxanthin to zeaxanthin diglucoside. Here, we report cloning of the crtX gene responsible for zeaxanthin diglucoside biosynthesis from Paracoccus haeundaensis and the production of the corresponding carotenoids in transformed cells carrying this gene. An expression plasmid containing the crtX gene (pSTCRT-X) was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 46 kDa. Biosynthesis of zeaxanthin diglucoside was obtained when the plasmid pSTCRT-X was co-transformed into E. coli containing the pET-44a(+)-CrtEBIYZ carrying crtE, crtB, crtI, crtY, and crtZ genes required for zeaxanthin $\beta$-D-diglucoside biosynthesis.

• Journal of Life Science
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• v.14 no.5
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• pp.752-760
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• 2004

The function of the [4Fe-4S] cluster containing iron (Fe-) protein in nitrogenase catalysis is to serve as the nucleotide-dependent electron donor to the MoFe protein which contains the sites for substrate binding and reduction. The ability of the Fe protein to function in this manner is dependent on its ability to adopt the appropriate conformation for productive interaction with the MoFe protein and on its ability to change redox potentials to provide the driving force required for electron transfer. The MgADP-bound (or off) conformational state of the nitrogenase Fe protein structure described reveals mechanisms for long-range communication from the nucleotide-binding sites to control affinity of association with the MoFe protein component. Two pathways, termed switches I and II, appear to be integral to this nucleotide signal transduction mechanism. In addition, the structure of the MgADP bound Fe protein provides the basis for the changes in the biophysical properties of the [4Fe-4S] observed when Fe protein binds nucleotides. The structures of the nitrogenase Fe protein with defined amino acid substitutions in the nucleotide dependent signal transduction pathways of the Switch I and Switch II have been determined by X-ray diffraction methods. These two pathways have been also implicated by site directed mutagenesis studies, structural analysis and analogies to other proteins that utilize similar nucleotide dependent signal transduction pathways. We have examined the validity of the assignment of these pathways in linking the signals generated by MgATP binding and hydrolysis to macromolecular complex formation and intermolecular electron transfer. The results provide a structural basis for the observed biophysical and biochemical properties of the Fe protein variants and interactions within the nitrogenase Fe protein-MoFe protein complex.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.27 no.2
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• pp.63-81
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• 1997

This study was performed in order to identify changes of the plasma membrane proteins in rat submandibular gland tumors induced by 7,12-dimethylbenz[a]anthracene [DMBA] and X-irradiation. Two kinds of tumor associated membrane proteins (protein A and B) were isolated with 3 M KCl extraction from rat submandibular gland tumors induced by DMBA and X-irradiation. To identify their antigenicities, immunoelectrophoresis and double immunodiffusion was carried out with various proteins extracted from liver, heart, skin and pancreas of adult rats and from embryonic liver, heart and skin. The rabbit antisera against the protein A did not cross-react with any of the proteins extracted from the above mentioned tissues, suggesting that protein A might be tumor specific antigen. However, the rabbit antisera against protein B was precipitated with proteins extracted from the liver of adult and embryonic rats. Polyacrylamide gel electrophoresis of these two proteins (A and B) showed that protein A was a dimer with molecular weights of 69,000 and 35,000 dalton, whereas protein B was a monomer with molecular weight of 50,000 dalton.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1116-1122
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• 2013

Cell line development is the most critical and also the most time-consuming step in the production of recombinant therapeutic proteins. In this regard, a variety of vector and cell engineering strategies have been developed for generating high-producing mammalian cells; however, the cell line engineering approach seems to show various results on different recombinant protein producer cells. In order to improve the secretory capacity of a recombinant tissue plasminogen activator (t-PA)-producing Chinese hamster ovary (CHO) cell line, we developed cell line engineering approaches based on the ceramide transfer protein (CERT) and X-box binding protein 1 (XBP1) genes. For this purpose, CERT S132A, a mutant form of CERT that is resistant to phosphorylation, and XBP1s were overexpressed in a recombinant t-PA-producing CHO cell line. Overexpression of CERT S132A increased the specific productivity of t-PA-producing CHO cells up to 35%. In contrast, the heterologous expression of XBP1s did not affect the t-PA expression rate. Our results suggest that CERT-S132A-based secretion engineering could be an effective strategy for enhancing recombinant t-PA production in CHO cells.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.139-146
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• 2006

The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.5
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• pp.551-556
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• 1996

Selenium (Se) apparent absorption and retention in sheep as influenced by diets differing in protein content through soybean meal supplementation was studied. A $3{\times}3$ Latin square design was used with three Japanese Corriedale wethers (45 kg average body weight), three periods, and three dietary treatments. In each period, 7 d dietary adjustment was followed by 5 d total collection of urine and feces. The three dietary treatments were : Diet 1, without soybean meal supplementation (14% crude protein, CP); Diet 2, with 10% soybean meal supplementation (16.5% CP); and Diet 3, with 20% soybean meal supplementation (19% CP). All the diets had a Se supplementation in the form of sodium selenite at 0.2 mg Se/kg dietary DM. The dietary DM intake of the animals was 2% of their body weight. No significant differences were obtained among the three dietary treatments of the Se balance of the animals. However, as percent of Se intake, only urinary Se concentration of Diet 3 was markedly lower (p < 0.05) than the other diets. Fecal Se as percent of Se intake followed the trend of Diet 3> Diet 2 > Diet 1 resulting a Se absorbed as percent of Se intake of 58.9%, 62.3% and 68.2% for Diets 3, 2 and 1, respectively but their differences among each other were insignificant. No significant differences that were observed either on Se retained as percent of intake (Diet 1, 48.2%; Diet 2, 45.2%; Diet 3, 46.0%) or Se retained as percent of Se absorbed (Diet 1, 70.7%; Diet 2, 72.4%; Diet 3, 77.9%). Significant correlation coefficients among the various measures of Se utilization were also observed. Regression analysis showed the following equation: Y = 93.8 - 1.86X (p <0.05, $r^{2}=0.48$), where Y is the Se absorbed as percent of Se intake (%) and X is the dietary protein content (%). This study concludes that Se requirement in sheep is greater when dietary protein content is high.