• Title/Summary/Keyword: MOPS

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Characterization and Developmental Regulation of Polysialyltransferase from Embryos of Strongylocentrotus nudus (둥근성게, Strongylocentrotus nudus 배에 존재하는 Polysialyltransferase의 특성 및 발현 조절에 관한 연구)

  • 남지흔;김영대;박영제;조진원
    • Development and Reproduction
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    • v.2 no.2
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    • pp.149-155
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    • 1998
  • The polysialic acid (polySia) glycotope covalently modifies cell surface glycoconjugates on cells as evolutionarily diverse as microbes and human. The recent chemical identification of polysialylated glycoproteins in the jelly coat and on the cell surface of the sea urchin egg raises important questions about their biosynthesis and possible function. Using CMP-[$^{14}$ C]Neu5Ac as substrate and cell free preparations from eggs and embryos of the sea urchin Stronglylcentrotus nudus, we have identified a membrane associated CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST) that transfers Neu5Ac to an endogenous acceptor. Optimal conditions for the polyST activity were found to be 2$0^{\circ}C$ in 20 mM MOPS buffer (pH 7.0). The polyST activity was increased 2.7 times by the addition of 10 mM $Mg^2$$^{+}$. The membrane-associated polyST also catalyzed the polysialylation of mammalian ganglioside GD3. Given that no structurally similar natural polysialylated gangliosides have been described, nor were observed in the present study, we conclude that a single polyST activity catalyzes sialylation of the endogenous acceptor and the gangliosides. Using an excess of GD3 as an exogenous acceptor, it was established that the expression of the polyST in S. nudus embryos increased rapidly at the mesenchyme blastula stage and reached at maximum at the gastrula stage. The finding that this polyST in the sea urchin embryo is developmentally regulated raises the possibility that it may play a role in the changing cell and tissue interactions that occur during gastrulation and the early stages of spicule formation.n.

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A simple screening method using lignoceullulose biodegradation for selecting effective breeding strains in Agaricus bisporus (리그노셀룰로오스 생물학적 분해를 이용한 간단한 양송이 육종효율 우수 균주 선발)

  • Oh, Youn-Lee;Nam, Youn-Keol;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.134-138
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    • 2017
  • The white button mushroom, Agaricus bisporus, is commercially the fifth most important edible mushroom, accounting for the production of 9,732 tons of mushrooms in Korea in 2015. The genus Agaricus has been known for its potential to degrade lignocellulosic materials. Chemical analyses carried out during the cultivation of A. bisporus indicated that the cellulose, hemicellulose, and lignin fractions were changed preferentially for both vegetative growth and sexual reproduction. We screened A. bisporus strains for effective biodegradation through extracellular enzyme activity using cellulase, xylanase, and ligninolytic enzymes. The enzyme biodegradations were conducted as follows: mycelia of collected strains were incubated in 0.5% CMC-MMP (malt-mops-peptone), 0.5 Xylan-MMP, and 0.5% lignin-MMP media for 14 days. Incubated mycelia were stained with 0.2% trypan blue. Eighteen strains were divided into 8 groups based on different extracellular enzyme activity in MMP media. These strains were then incubated in sterilized compost and compost media for 20 days to identify correlations between mycelial growth in compost media and extracellular enzyme activity. In this study, the coefficient of determination was the highest between mycelial growth in compost media and ligninolytic enzyme activity. It is suggested that comparison with ligninolytic enzyme activity of the tested strains is a simple method of screening for rapid mycelial growth in compost to select good mother strains for the breeding of A. bisporus.

Cellular Iron Uptake from Aqueous Solutions depending on Reaction Conditions by genetically engineered Saccharomyces cerevisiae (재조합 Saccharomyces cerevisiae에 있어서 반응조건에 따른 수용성 철의 생체 흡수)

  • Kim Sang-Jun;Chang Yu-Jung;Park Chung-Ung;Jeong Yong-Seob;Kim Kyung-Suk
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.441-445
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    • 2004
  • Cellular iron uptake was performed in the yeast Saccharomyces cerevisiae that transformed with human ferritin H- and L-chain genes. The recombinant yeasts were enriched in YEP medium supplemented with $2\%$ galactose for 3 days and the iron uptake was followed by incubating the cells with iron in 20 mM MOPS buffer (pH 6.5). The reactions were examined under different conditions including the iron compounds of Fe(II) and Fe(III), the concentration of iron, the concentration of cells and the reaction time. From our results, the recombinant yeast YGH2 producing H-chain ferritin showed higher cellular iron concentration at the cell concentration of 100 mg/ml than 200 mg/ml. Iron presented as Fe(II) rather than Fe(III) was taken up more efficiently. Iron uptake increased slightly when iron was added up to 14.3 mM Fe(II) and then its cellular iron concentration was $16.7{\pm}0.7\;{\mu}mol/g$ cell wet wt. In addition, the iron uptake reaction reached to maximum at about 2 hr incubation.

Research on Pilot Decision Model for the Fast-Time Simulation of UAS Operation (무인항공기 운항의 배속 시뮬레이션을 위한 조종사 의사결정 모델 연구)

  • Park, Seung-Hyun;Lee, Hyeonwoong;Lee, Hak-Tae
    • Journal of Advanced Navigation Technology
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    • v.25 no.1
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    • pp.1-7
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    • 2021
  • Detect and avoid (DAA) system, which is essential for the operation of UAS, detects intruding aircraft and offers the ranges of turn and climb/descent maneuver that are required to avoid the intruder. This paper uses detect and avoid alerting logic for unmanned systems (DAIDALUS) developed at NASA as a DAA algorithm. Since DAIDALUS offers ranges of avoidance maneuvers, the actual avoidance maneuver must be decided by the UAS pilot as well as the timing and method of returning to the original route. It can be readily used in real-time human-in-the-loop (HiTL) simulations where a human pilot is making the decision, but a pilot decision model is required in fast-time simulations that proceed without human pilot intervention. This paper proposes a pilot decision model that maneuvers the aircraft based on the DAIDALUS avoidance maneuver range. A series of tests were conducted using test vectors from radio technical commission for aeronautics (RTCA) minimum operational performance standards (MOPS). The alert levels differed by the types of encounters, but loss of well clear (LoWC) was avoided. This model will be useful in fast-time simulation of high-volume traffic involving UAS.

Effect of culture pH and media composition on the ratio of tcicoplanin $A_1$ and $A_2$ biosynthesis

  • Kim, Yun-Jeong;Song, Yun-Seok;No, Yong-Taek
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.325-328
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    • 2001
  • Teicoplanin is a glycopeptide antibiotic produced by Actinoplanes teichomyceticus novo sp. A TCC 31121. It is active against Gram-positive bacteria and it is under evaluation for use in man. Teicoplanin mixture in fermentation broth contains major amounts of teicoplanin $A_1$ and $A_2$ and a minor amount tcicoplanin of $A_3$. Commercial teicoplanin product is composed of five major components of very similar polarity, designated T-$A_2$-l, 2, 3, 4 and 5, and the more polor component, designated T -$A_3$. The culture conditions were studied in order that hydrophilic teicoplanin $A_2$ components are more produced but hydrophobic teicoplanin $A_1$ with lower bioactivity are less produced in submerged culture. Effects of culture pH and nutrients on the biosynthes ratio of teicoplanin $A_1$ and $A_2$ were confirmed in flask culture using MOPS buffer system through TLC, bioautography and bioassay. It was elucidated that optimal pH is 7.4 for teicoplanin $A_2$ biosynthesis but is 5.2 for teicoplanin $A_1$ biosynthesis, and that trace elements stimulate $A_2$ production but malt extract stimulate $A_1$production.

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A Process of the Technical Performance Management for A Space Launch Vehicle R&D Project (우주발사체 개발사업을 위한 기술성능관리 프로세스)

  • Yoo, Il Sang;Cho, Dong Hyun;Kim, Keun Taek
    • Journal of the Korean Society of Systems Engineering
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    • v.10 no.2
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    • pp.71-79
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    • 2014
  • To enhance success probability of a system development project, its overall risk level should be minimized through systematically managing schedules, costs, and technical performances. However, Attempts to manage technical performance compared to numerous efforts to control costs and schedules in such projects are deficient. Particularly, a space launch vehicle, a large complex system, development project is much less likely to meet its technical performance objectives due to its technological difficulty, along with schedule delay and cost overrun. The technical performance management (TPM) is a method for tracking and managing technical progress in order to achieve technical performance targets within schedule and budget. In this paper, we investigate applications of the TPM in several space launch vehicle development projects. Then we propose and validate the TPM process to achieve a successful mission in such projects.

Reaction between Gas-phase Hydrogen Atom and Chemisorbed Bromine Atoms on a Silicon(001)-(2X1) Surface

  • Park, Jong-Keun;Ree, Jong-Baik;Lee, Sang-Kwon;Kim, Yoo-Hang
    • Bulletin of the Korean Chemical Society
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    • v.28 no.12
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    • pp.2271-2278
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    • 2007
  • Electron transfer of a redox protein at a bare gold electrode is too slow to observe the redox peaks. A novel Nafion-riboflavin functional membrane was constructed during this study and electron transfer of cytochrome c, superoxide dismutase, and hemoglobin were carried out on the functional membrane-modified gold electrode with good stability and repeatability. The immobilized protein-modified electrodes showed quasireversible electrochemical redox behaviors with formal potentials of 0.150, 0.175, and 0.202 V versus Ag/AgCl for the cytochrome c, superoxide dismutase and hemoglobin, respectively. Whole experiment was carried out in the 50 mM MOPS buffer solution with pH 6.0 at 25 oC. For the immobilized protein, the cathodic transfer coefficients were 0.67, 0.68 and 0.67 and electron transfer-rate constants were evaluated to be 2.25, 2.23 and 2.5 s?1, respectively. Hydrogen peroxide concentration was measured by the peroxidase activity of hemoglobin and our experiment revealed that the enzyme was fully functional while immobilized on the Nafion-riboflavin membrane.

Characterization of the Catalytic Properties of Recombinant Acetohydroxyacid Synthase from Tobacco

  • Kim, Joung-Mok;Choi, Jung-Do;Kim, Bok-Hwan;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • v.26 no.2
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    • pp.260-264
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    • 2005
  • The nature of the active site of Tobacco acetohydroxyacid synthase (AHAS) in the substrate- and cofactorbinding was studied by kinetics and fluorescence spectroscopy. The substrate saturation curve does not follow Michaelis-Menten kinetics at different temperatures (7, 21 and 37 ${^{\circ}C}$), pH (6.5, 7.5 and 8.5) and buffers (Tris-HCl and MOPS). The concentration of one half of the maximum velocity ($S_{0.5}$) decreased in the following order: pyruvate $\gt$ ThDP $\approx$$Mg^{+2}$ $\gt$ FAD. However, the catalytic efficiency (K$_{cat}/S_{0.5}$) inversely decreased in the following order; FAD $\gt$ $Mg^{+2}$ $\approx$ThDP $\gt$ pyruvate, indicating that the cofactors by in decreasing order; FAD, $Mg^{+2}$, ThDP, affect the catalysis of AHAS. The dissociation constant ($K_d$) of the intrinsic tryptophan fluorescence decreased with the same tendency of the concentration of one half of the maximum velocity ($S_{0.5}$) decreasing order. This data provides evidence that the substrate and cofactor binding natures of the active site, as well as its activation characteristics, resemble those of other ThDP-dependent enzymes.

Biochemical Characterization of 1-Aminocyclopropane-1-Carboxylate Oxidase in Mung Bean Hypocotyls

  • Jin, Eon-Seon;Lee, Jae-Hyeok;Kim, Woo-Taek
    • BMB Reports
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    • v.31 no.1
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    • pp.70-76
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    • 1998
  • The final step in ethylene biosynthesis is catalyzed by the enzyme 1-aminocyclopropane-1-carboxylate (ACC) oxidase. ACC oxidase was extracted from mung bean hypocotyls and its biochemical characteristics were determined. In vitro ACC oxidase activity required ascorbate and $Fe^{2+}$, and was enhanced by sodium bicarbonate. Maximum specific activity (approximately 20 nl ethylene $h^{-1}$ mg $protein^{-1}$) was obtained in an assay medium containing 100 mM MOPS (pH 7.5), $25\;{\mu}M$ $FeSO_4$, 6 mM sodium ascorbate, 1 mM ACC, 5 mM sodium bicarbonate and 10% glycerol. The apparent $K_m$ for ACC was $80{\pm}3\;{\mu}M$. Pretreating mung bean hypocotyls with ethylene increased in vitro ACC oxidase activity twofold. ACC oxidase activity was strongly inhibited by metal ions such as $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, and $Mn^{2+}$, and by salicylic acid. Inactivation of ACC oxidase by salicylic acid could be overcome by increasing the $Fe^{2+}$ concentration of the assay medium. The possible mode of inhibition of ACC oxidase activity by salicylic acid is discussed.

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Isolation and Characterization of Phosphorus Accumulating Microorganisms under Liquid Fertilization of Swine Slurry (액비화 과정 중 인 이용 우수미생물 분리 및 특성)

  • Lim, Joung-Soo;Cho, Sung-Back;Hwang, Ok-Hwa;Yang, Seung-Hak
    • Journal of Animal Environmental Science
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    • v.20 no.2
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    • pp.77-84
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    • 2014
  • This study was conducted to investigate the bacterial capability to accumulate phosphorus during liquid composting process of pig slurry. Storage liquid compost and pig slurry were analyzed by using MALDI-TOF technique, which showed the colonies of Acinetobacter towneri and Bacillus licheniformis. In addition, bacterial colonies were isolated under high phosphoric acid conditions using X-phosphate MOPS medium with the addition of 2 mM $K_2HPO_4$. Microbial growth was observed in high and low phosphoric conditions due to the growth of bacterial diversities in the liquid fertilizer and slurry. The colonies isolated in the high phosphoric acid medium were uncultured bacterium clone and Acinetobacter sp. were identified by analysis of 16S rRNA gene sequences. Uncultured bacterium showed higher growth rate and excellent phosphorus ability then Acinetobacter sp.. In addition to Paenibacillus sp. AEY-1 isolated from pig slurry performed excellent phosphorus utilizing capability.