• Journal of Embryo Transfer
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• v.25 no.4
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.177-188
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• 1996

The effects of cytokines on the growth and differentiation of glial cells in culture were evaluated to confirm that cytokines could modify the number and function of glial cells. Proliferation of glial cells was determined by the $^3H-thymidine$ uptake and the double immunostain with anti-cell specific marker and anti-bromodeoxyuridine(BrdU) antibody. To check the effect on the differentiation of glial cells, the amount of glial fibrillar acidic protein(GFAP) and the activity of glutamine synthetase(GS) were measured in astrocytes. And also the amounts of myelin basic protein(MBP) and the activity of 2',3'-cyclic nucleotide phosphohydrolase(CNPase) were measured in oligodendrocytes. Among the cytokines used, only interleukin-$1{\beta}(IL-1{\beta})$ stimulated the growth of type 1 and type 2 astrocyte as well as 0-2A precursor cell. When the functional changes in these glial cells by cytokines were tested, $IL-1{\beta}$ did not increase GFAP content in type 1 and type 2 astrocyte, but $IL-1{\beta}$ increased GS activity in type 1 astrocyte, and slightly decreased this enzyme activity in type 2 astrocyte. Also interleukin-2(IL-2) and $interferon-{\gamma}$ $(IFN-{\gamma})$ inhibited the activity of GS in type 1 and type 2 astrocyte. On the other hand, all cytokines used did not modify the growth and differentiation in oligodendrocytes. From these results we could suggest that $IL-1{\beta}$ increases the growth of type 1 and type 2 astrocyte and also promotes the development for 0-2A precursor cell to type 2 astrocyte.

• Journal of Life Science
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• v.20 no.1
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• pp.97-102
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• 2010

Streptomyces platensis YK-2, newly isolated from forest soil, produces transglutaminase (TGase), which catalyses an acyl transfer reaction between the primary grade amine and protein or $\gamma$-carboxyamide group of peptide bound glutamine residues. For a molecular genetic study of S. platensis, an effective transformation method was established by using a conjugal transfer of DNA from Escherichia coli to spores of actinomycetes. The highest transconjugation frequency of S. platensis was obtained on an MS medium containing 50 mM $MgCl_2$, using $5{\times}10^7\;E$. coli as a DNA donor and $1{\times}10^8$ spores without heat treatment as a host. We also identified that S. platensis contains a single attB site within an ORF encoding a pirin-homolog, and that its attB site sequence shows high homology to that of S. logisporoflavus. In addition, it was confirmed by phenotypic analyses of exconjugants that the introduction of heterologous DNA into the attB site of the S. platensis chromosome does not affect its morphological differentiation and TGase production.

• Journal of Plant Biology
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• v.4 no.1
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• pp.9-11
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• 1961

A comparative investigation of free amino acids content in healthy check and virus diseased leaves of Chinese date tree, Zyzyphus jujuqa Mill var. inermis Rhed, was carried out by authors throughout the growing season of 1959 and 1960 from June to October. The methods of qualitative analysis of free amino acids aplied in this experiment is followed by Moore and Stein. Free amino acids determined in this experiment are shown in Fig. 1 and Table 1. As the figure and the table are shown, three more amino acids such as glutamine, asparagine and histidine are detected in the diseased material. The additional amino acids which are known as diamines in diseased leaves are conspicuous. It is presumed that the diamine might be incresed by the self-reproduction of the virus in cooporation with certain enzymes which are carrying out the protein metabolism in the host protoplast in contrast with the healthy checks which is carrying out normal protein metabolism. From the histological poing of view, the facts of phloem degeneration or necrosis in diseased leaves, it seems to interrupt to move free amino acids from roots to leaves and it possibly takes place an excessive production of NH3 which is diaminated by the metabolism of nitrogen compounds in such conditioned leaves. Therefore, it is also presumed that additional diamino acids are accumulated in diseased leaves. There are no change of amino acids are accumulated in diseased leaves. There are no change of amino acids in both materials of this plant throughout the growing season qualitatively, and this result agress with the paper of Knight.

• Journal of Life Science
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• v.22 no.11
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• pp.1451-1455
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• 2012

The study was conducted to investigate the effect of Maillard reaction products (MRPs) on enzymatic browning of crown daisy (Chrysanthmum coronarium var. spatiosum). The MRPs prepared by heating various amino acid and sugar at $90^{\circ}C$ caused a strong inhibitory effect on crown daisy polyphenol oxidase (PPO, ${\sigma}$-diphenol oxygen oxidoreductase, EC 1.10.3.1). As the reaction time of the solution containing glycine and glucose increased at $90^{\circ}C$, the production of MRPs was increased, whereas the amounts of glycine and glucose were decreased. Accordingly, the inhibitory effect of crown daisy PPO activity by MRPs was increased as the amounts of synthesized MRPs were increased. The MRPs synthesized from the various amino acids and sugars significantly reduced the PPO activity, particularly MRPs prepared by glutamine and xylose. The Michealis-Menten constant value ($K_m$) of crown daisy PPO with catechol as a substrate was 22.0 mM, and MRPs were a noncompetitive inhibitor against crown daisy PPO.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.272-280
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• 1991

The effect of media feeding rate on cell growth and monoclonal antibody production in the fed-batch culture ot hybridoma A4W was studied. In the batch culture, the highest specific antibody production rate was observed at the begining of the culture period but its value tended to decrease rapidly with the culture time. The final antibody concentration and volumetric productivity was 65 $\mu g$/ml and 13 mg Mab/l/day, respectively. In the fed-batch culture, the specific antibody production rate, $q_p$ rebounded sharply within a few hours after the media feeding was started and it remained high until the end of culture if the media feeding was continued. The final antibody concentration was 220 $\mu g$/ml and the volumetric productivity was 45.1 mg/l/day. Further increase in final antibody concentration was achieved by applying a modified media of which component was fortified with glucose and glutamine, hence the final antibody concentration in this case was 270 $\mu g$/ml and the volumetric productivity was 51.8 mg/lday, which is as four tinlcs as high cuixparinf! to that of batch culture.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.273-277
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• 2004

$\alpha$-L-Arabinofuranosidase ($\alpha$-L-AFase, EC 3.2.1.55) was isolated from hyperthermophilic microorganism, Thermotoga maritima. The open reading frame (ORF) of $\alpha$-L-AFase gene is 1,455 bp long and encodes 484 amino acid residues with a molecular weight of 55,265 Da. The ORF of $\alpha$-L-AFase gene was introduced into the E. coli expression vector, $_p/RSET-B, and overexpressed in E. coli BL21. The purified recombinant $\alpha$-L-AFase showed the highest activity at 10$0^{\circ}C$ and pH 5.5. The purified enzyme appeared to have no metal cofactor requirement. The Km and specific activity values of the recombinant enzyme were 0.99 mM and 1,200 U/mg on p-nitrophenyl-$\alpha$-L-arabinofuranoside. It released only L-arabinose from sugar beet arabinan, sugar beet debranched arabinan and oat spelts arabinoxylan but had no activity onarabinogalactan and gum arabic. This result suggests that L-arabinose could be produced from natural polysaccharides using this enzyme. Mutant enzymes which Glu26, Glu172 and Glu281 residues were replaced to alanine, aspartic acid or glutamine caused Kcat to decrease by a factor of between 10$^3$ and 10$^4$. Glu172 and Glu281 residues of $\alpha$-L-AFase are seemed to be the acid/base and nucleophile in catalytic reaction, respectively, and Glu26 is supposed to playa key role in substrate binding.ng.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.757-761
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• 2002

Proximate composition, phenolic compounds, and amino acid composition of molokhia (Corchorus olitorius) and molecular weight distribution of its mucilage were determined. Concentrations of dietary fibers and protein were 37.4 and 24.4% of the total dry weight, respectively. Mucilage was obtained by extracting dry molokhia leaves with hot water, followed by the addition of 55% ethanol. Although about 50% of the mucilage was lost during the extration, it was still rich in polyphenol compounds. In mucilage, proteins were composed of glutamic acid, glycine, alanin, and aspartic acid, and molecular weight of most polysaccarides ranged from $40,000{\sim}500,000$ daltons based on gel filtration chromatography.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.1-9
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• 1998

This study was designed to determine the most suitable manufacturing conditions of stewed pumpkin juice(SPJ) in terms of nutrition and taste. Well ripened pumpkin was heated for 3, 4, 5, 6 and 7 hours in a pressure cooker and then extracted and packed in retort pouches. These were kept at 4$^{\circ}C$ and 28$^{\circ}C$ for 30 days, respectively and changes of the physicochemical components of SPJ were investigated. The free amino acid compositions of pumpkin and SPJ occupied asparagine, aspartic acid, glutamine, arginine and serine in increasing order of abundance. Except SPJ heated for 3 hours, SPJ stored at 28$^{\circ}C$ had the most abundant free amino acids, followed by SPJ stored at 4$^{\circ}C$, and SPJ not stored. The fatty acid composition of pumpkin occupied behenic acid, erucic acid, palmitic acid, linoleic acid, heneicosanoic aicd, in order of abundance. When SPJ was heated over 3 hours, palmitic acid and linolenic acid decreased greatly. SPJ stored at 4$^{\circ}C$ and 28$^{\circ}C$ increased contents of saturated fatty acids such as behenic acid, palmitic acid than SPJ not stored. As SPJ was heated for a long time, carotenoid was mostly destroyed and the Hunter "L", "a" and "b" values of SPJ decreased. Among the mineral constituents of SPJ, Na and Ca were dominantly occupied. It was found that the range of the total visible cell count was 3~4CFU/ml from the SPJ which was heated for 3 hours and it increased by 1$\times$109CFU/ml when the SPJ was stored at 28$^{\circ}C$. It seems that the contamination by microorganism have occurred at packing process. In sensory evaluation, the SPJ which was heated for 5 hours had the highest scores in overall preference, sweet smell and sweetness.ce, sweet smell and sweetness.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.1
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• pp.225-232
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• 1999

Apo E polymorphism(e2, e3, e4) was among the first reported genetic polymorphism that explained part of the normal variation in plasma cholesterol concentrations. Among 62 normolipidemic healthy females, aged 19 up to 22 years, the relative frequencies of E3/3 was 0.806(n=50), E3/2 was 0.081(n=5), E3/4 allele was 0.113(n=7), and no E2/2, E2/4 and E4/4 were found. Based on the five samples of E2 allele, five subjects were randomly selected by E3 and E4 groups for the study of effects of apo E polymorphism on the distribution of serum lipid and amino acids profiles. No differences in the anthro pometric data among apo E isomers were found, otherwise the pulsation was higher in E4 than that in the others. There were no differences in plasma total HDL , HDL3 , HDL2 & LDL cholesterol, and apo A I concentrations. However, phenotype means significantly rank E2>E3>E4 allele in average TG levels(p=0.014), and rank E4>E3>E2 in total cholesterol levels(p=0.011). Atherogenic index(AI) such as lipoproteins was significantly increased in E2 & E4 than that in E3(p=0.045). Subjects with E3/2 allele had significantly higher concentrations of glutamine, phosphoserine and taurine, while subjects with E3/4 allele showed significantly lower concentrations of arginine and am inobutyrate and elevated level of phosphoserine in plasma com pared to those of E3/3 allele. Higher level of plasma taurine in subjects with E3/2 or E3/4 allele appears to be related to the elevated level of plasma total and LDL cholesterol concentrations compared to those of E3/3 allele.