• Title/Summary/Keyword: LEU

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Comparision of PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS) in Leu72Met Polymorphism of Ghrelin Gene (Ghrelin 유전자의 Leu72Met 다형성 분석에서 PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS)의 비교분석)

  • Kang, Ju Sung;Kim, Se Rim;Kim, Sun Young;Joo, Chan Uhng;Cho, Soo Chul;Hwang, Pyoung Han
    • Clinical and Experimental Pediatrics
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    • v.48 no.10
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    • pp.1068-1075
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    • 2005
  • Purpose : The role of ghrelin, which promotes the secretion of growth hormone, was not well known until now. Recently it was found that the mutation of ghrelin gene is related to obesity and diabetes. This study is to find the screening method that can easily and effectively detect the polymorphism of Leu72Met in ghrelin gene of obesity patients and apply it to clinical usage. Methods : We compared PCR-RFLP, PCR-SSCP and ARMS methodologies for analyzing of the polymorphism of Leu72Met in ghrelin gene of obesity children, and also studied the merits and demerits of these methodologies. Results : In this study, we were able to find out the band of peculiar allele of Leu72Met in ghrelin gene using PCR-RFLP, PCR-SSCP and ARMS analyses. The polymorphism of Leu72Met in ghrelin gene determined by all above methodologies was in complete agreement. Compared to the PCR-RFLP and PCR-SSCP, ARMS analysis is simple, inexpensive and also consume less time. It is very sensitive to analyze the polymorphism and easy to understand the results of test. Conclusion : Though PCR-RFLP, PCR-SSCP and ARMS analyses were sensitive to analyze the polymorphism of Leu72Met in ghrelin gene, ARMS analysis appears to be more efficient than PCR-RFLP and PCR-SSCP. Therefore, we conclude that ARMS analysis is suitable to analyze the polymorphism of Leu72Met in ghrelin gene for large quantity of specimens.

Synthesis and Functional Properties of Plastein from the Enzymatic Hydrolysates of Filefish Protein. 3. Functional Properties of Plasteins (말쥐치육 단백질의 효소적 가수분해물을 이용한 Plastein의 합성 및 그 물성 , 3. Plastein의 기능성)

  • KIM Se-Kwon;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.6
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    • pp.582-590
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    • 1987
  • Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper. L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. The structural changes of freeze-dried filefish meat, peptic hydrolysate, FPC and plasteins were observed by Scanning Electron Microscopy(SEM). The functional properties of plasteins also were measured. The solubility of plasteins was higher than that of FPC and the Glu-plastein had $95\%$ solubility in the range of pH 3-10. The dispersibility of Glu-plastein and protease plastein was similar to that of egg albumin, but those of the other plasteins were lower. The water holding capacity of plasteins was lower than that of egg albumin and C. Lipid absorption of Leu-plastein was tile highest, holding 1.80 ml/g, and that of the other plasteins was similar to that of egg albumin. The emulsifying activity of Leu-plastein was the highest, holding $61.2\%$, and that of Glu-plastein was the lowest, holding $50.7\%$. The emulsifying stability of plasteins was similar to that of the emulsifying activity. The emulsifying capacity of Leu-plastein was 384 ml/g(the highest), but that of Glu-plastein and $\alpha-chymotrypsin$ plastein was 248 ml/g(the lowest). The Leu-plastein shelved the highest foaming capacity, $373\%$. The foaming capacity of other plasteins was higher than that of egg albumin. The foaming stability of plasteins was superior to that of egg albumin. The viscosity of plasteins was lower than that of egg albumin. The microstructure of $\alpha-chymotrypsin$ plastein by SEM wassimilar to that of papain plastein, but other plasteins showed differences in their microstructure. The microstructure of Glu-plastein had a smooth shape.

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Synthesis and In-vitro Evaluation of N4-Amino Acid Derivatives of Cytarabine for Improving the Oral Delivery of Cytarabine

  • Jin, Ming-Ji;Hong, Joon-Hee;Han, Hyo-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.38 no.4
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    • pp.255-259
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    • 2008
  • The present study aimed to investigate the in-vitro characteristics of N4-amino acid derivatives of cytarabine for the oral delivery of cytarabine. After the synthesis of L-Ile-cytarabine, L-Leu-cytarabine and L-Arg-cytarabine, the gastrointestinal stability of each prodrug was examined using artificial gastric juice and intestinal fluids. The cellular uptake characteristics of prodrugs were also examined in Caco-2 cells. While L-Ile-cytarabine and L-Leu-cytarabine appeared to be stable in all the tested biological media during 4-hr incubation, L-Arg-cytarabine was rapidly disappeared within 5 min. Accordingly, the cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine was significantly higher than that of its parent drug, cytarabine in Caco-2 cells but the cellular uptake of L-Arg-cytarabine was similar to that from its parent drug. The cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine appeared to be saturable as drug concentration increased from 0.4 to 4 mM. Collectively, L-Ile-cytarabine and L-Leu-cytarabine could be promising candidates to improve the oral absorption of cytarabine via a saturable transport pathway.

Purification and Characterization of the Anabolic Acetolactate Synthase III from Serratia marcescens ATCC 25419

  • Joo, Han-Seung;Kim, Soung-Soo
    • BMB Reports
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    • v.34 no.3
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    • pp.244-249
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    • 2001
  • The anabolic acetolactate synthase III was purified to homogeneity from Serratia marcescens using DEAE-Sepharose, Phenyl-Sepharose, and hydroxylapatite column chromatography The native molecular weight of the enzyme was approximately 165 kDa. The enzyme is composed of two large and two small subunits with molecular weights of 64 and 15 kDa, respectively. The N-terminal sequence of the large and small subunit of the enzyme was Ser-Ala-Thr-Pro-Gln-Pro-Ser-Thr-Arg-Phe-Thr-Cys-Ala-Gln-Leu-Ile-Ala-His-Leu and Met-Leu-Gln-Pro-Gln-Asp-Lys-Pro-Gln-Val-Ile-Leu-Glu-Leu-Ala-Val-Arg-Asn-His-Pro-Gly-Val-Met-Ser-His-Val, respectively. The optimum pH and pI value were 7.5 and 5.5, respectively The $IC_{50}$ values were $20\;{\mu}M$ and $14\;{\mu}M$ for valine and herbicide SU7, respectively. The substrate specificity ratio, R value, was determined to be approximately 40, which suggests that this enzyme prefers the formation of $\alpha$-aceto-$\alpha$-hydroxybutyrate leading to the synthesis of isoleucine.

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Molecular Cloning of the Gene Coding for 3-Isopropylmalate Dehydrogenase of Kluyveromyces fragilis (Kluyveromyces fragilis의 LEU gene의 Cloning)

  • 박성희;이동선;우주형;김종국;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.305-308
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    • 1990
  • In order to clone the gene coding for 3-isopropylmalate dehydrogenase of Muyveromyces fragilis, a shuttle plasmid vector pHNll4 was used. It can serve as a cloning vector in Saccharomyces cerevisiae DBY746 for other Sau3AI-cleaved DNA segment of Kluyveromyces fragilis. Two cloned fragments which complement the leu2 mutation of Saccharomyces cerevisiae and E, coli were obtained. Their length was 4.4 kb an 3.5 kb, and their orientation was opposite each other. From the fact that the two recombinant plasmids were expressed in Saccharomyces cerevisiae and E, coli, probably the two inserts had the promoter of Ktuyveromyces fi-agilis and that of Kluyveromyces fiagilis was efficiently assosiated with RNA polymerase of Saccharomyces cerevisiae and E. coli. According to the result of Southern hybridization, we thought that the cloned fragment has low homology with 3-isopropylmalate dehydrogenase coding region of E. coli and Saccharomyces cerevisiae.

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Improvement of Bacterial Endo-1,4-,\beta-D-glucanase(CMCase) Secretion in Yeast by Mutagenesis of Glucoamylase Signal Sequence. (Glucoamylase 분비신호서열의 돌연변이에 의한 효모에서 세균의 Endo-1,4-\beta-D-glucanase의 분비능 증진)

  • 이준원;강대욱;김보연;오원근;민태익;이상원;변유량;안종석
    • Microbiology and Biotechnology Letters
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    • v.28 no.4
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    • pp.195-201
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    • 2000
  • Glucoamylase of Saccharomyces diastaticus is produced as a large precursor composed of signal peptide (21 amino acid residues), Thr and Ser-rich region and functional glucoamylase. To evaluate the utility of the glucoamylase signal peptide (GSP) for the secretion of foreign proteins, four types of GSP mutants (ml : Pro-18 longrightarrowLeu-18, m2 : Tyr-13 longrightarrowLeu, m3 : Ser-9longrightarrowLeu-9, m4 : Asn-5 longrightarrowPro-5) were constructed and secretion efficiency of each mutant was compared with that of native GSP by the expression and secretion of Bacillus subtilis CMCase under the control of GAP in N-terminal domain and hydrophobic domain. n mutant 4, a polar amino acid was replaced by a helix - breaking Pro residue. CMCase activity assay and Western blot analysis revealed that CMCase secretion by GSP mutants replaced by Leu were increased compared with native GSP. In the case of m2 and m3, the substitution of Leu for Tyr-13 and Ser-9 in the hydrophobic region resulted in a twofold increase in the extracellular CMCase activity.

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Chemical Constituents of Cynochum wilfordi , Codonopsis lanceolata and lxeris sonchifolia (백하수오 , 더덕 , 고들빼기의 화학성분)

  • 이상래
    • Korean Journal of Plant Resources
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    • v.5 no.1
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    • pp.25-29
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    • 1992
  • The proximate and amino acid composition of wild and cultivated Cynochum wilfordi, Codopsis lancolata and lxeris sonchifolia were analyzed. Cynochum wilfordi, had higher contents of sugar ascompared with Codonopsis lancolata and lxeris sonchifolia. Fats was 2-3 times higher in Cynochumwilordi then Codonopsis lancolata and lxeris sonchifolia. No significant difference in the contents wasfound between the open field root, spontanity and cultivated at 30'c in biotronroom at condonopsislanceolata. However, contents of protein was 12.8% in lxeris sonchifolia. Total amino acid were 1.5-1.8 times higher in lxeris sonchifolia. than Cynochum wilfordi and Codonopsis lancolata. The decresingorder of the amounts of amino acid infused from the Cynochum wilfordi samples was Glu, Arg, Asp,Leu, Pro, Lys, Codonopsis lancolata (Changdeuk) was Arg, Glu, Pro, Leu, Asp, Lys, Codonopsislanceilata(Chirisan) was Arg, Glu, Pro, Asp, Lys, Leu, Codonopsis lancolata(Yeanbeun) was Arg,Glu, Asp, Leu, Ser, Lys, and lxeris sonchifolia was Glu, Arg, Asp, Cys, Lys, Pro.

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Characterization of an Antibiotic Produced by Bacillus subtilis JW-1 that Suppresses Ralstonia solanacearum

  • Kwon, Jae Won;Kim, Shin Duk
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.13-18
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    • 2014
  • Bacillus subtilis JW-1 was isolated from rhizosphere soil as a potential biocontrol agent of bacterial wilt caused by Ralstonia solanacearum. Seed treatment followed by a soil drench application with this strain resulted in >80% reduction in bacterial wilt disease compared with that in the untreated control under greenhouse conditions. The antibacterial compound produced by strain JW-1 was purified by bioactivity-guided fractionation. Based on mass spectroscopy and nuclear magnetic resonance spectral data ($^1H$, $^{13}C$, $^1H-^1H$ correlation spectroscopies, rotating frame nuclear Overhauser effect spectroscopy, and heteronuclear multiple-bond correlation spectroscopy), the structure of this compound was elucidated as a cyclic lipopeptide composed of a heptapeptide (Gln-Leu-Leu-Val-Asp-Leu-Leu) bonded to a ${\beta}$-hydroxy-iso-hexadecanoic acid arranged in a lactone ring system.

Screening of high Antibacterial Lactic Acid bacteria for the Preparation of Dongchimi-Juice for Naengmyon (냉면용 동치미액 제조를 위한 항균 활성이 높은 젖산균의 선발)

  • 조신호
    • The Korean Journal of Food And Nutrition
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    • v.12 no.1
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    • pp.69-76
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    • 1999
  • In order to screen lactic acid bacterial starter having high antibacterial activity and giving good flavor in Dongchmi for Naengmyon antibacterial activity and acid producing ability of 24 strains of Kimchi lactic acid bacteria were tested. Among 24 strains tested Leuconostoc mesenteroides subsp. mesenteroides C16, Leu. paramesenteroides C18 Lactobacillus bavaricus B01, B06, C19, C32 Lac. homohiochii B21 and B22 showed high antibacterial activity and their antibacterial activities were more active against Listeria monocytogenes and Staphylococcus aureus than against Escherichia coli and Salmonella typhimurium. The odors of Dongchimi-juices fermented with Leu. mesenteroides or Leu. paramesenteroides were more favorable than those with Lac. bavaricus or Lac. homohiochii. But the odors of Naengmyon-broths which were made with each Dongchimi-juice were not significantly different each other. In consideration of antibacterial ac-tivity and flavor of Dongchimi-juice two strains of lactic acid bacteria Lac. homohiochii B21 and Leu. mes-enteroides subsp. mesenteroides C16 were finally screened out as Dongchimi starters.

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Prompt neutron lifetime calculations for the NIRR-1 reactor

  • Ibrahim, Yakubu V.;Adeleye, Micheal O.;Njinga, Raymond L.;Odoi, Henry C.;Jonah, Sunday A.
    • Advances in Energy Research
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    • v.3 no.2
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    • pp.125-131
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    • 2015
  • Prompt neutron lifetime calculations have been performed for the NIRR-1 reactor HEU and LEU cores using the 1/v insertion and the Adjoint flux weighing methods. Results of calculations obtained for the HEU and LEU cores are respectively $57.3{\pm}0.8$ and $47.5{\pm}0.7$ for the 1/v insertion and $56.9{\pm}0.3$ and $46.3{\pm}0.5$ for the Adjoint flux. There is a good agreement seen between the two methods for both cores. The prompt neutron lifetime was observed to be shorter in the LEU than for the HEU as expected. However, the Adjoint flux weighing method seemed to be the easiest method in calculating the prompt neutron lifetime for NIRR-1.