• Title/Summary/Keyword: NADH

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Identification of mitochondrial mutant (NADH-dehydrogenase) using PCR method and regeneration of mutants from Zea mays (PCR 기법을 사용한 옥수수 미토콘드리아 변이체 (NADH-dehydrogenase)의 선별과 재분화)

  • 설인환
    • Journal of Life Science
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    • v.8 no.1
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    • pp.8-13
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    • 1998
  • The maize mitochondrial mutant (NCS2) is derived from homologous recombination between genes encoding NADH dehydrogenase subunit 4 and subunit 6. Plants from mitochondria mutants exhibited severe related growth and development including dwarfism and striping on the leaves. Aborted embryos from NCS2 mutants have been rescued and cultured on the N6 medium supplemented with 2,4-D 1 mg/l. Most calli from NCS2 aborted embryos showed slow growing pattern at first stage. However, upon continuous culturing them on the medium, those were segregated into mutant and normal callus lines. These segregations could be detected by using PCR method with three primers. Such segregation seems to be resulted from the preferential growth of normal cells over the mutant cells on the normal culture condition. Therefore, this method can be used for determining rate of indirect cytoplasmic segregation by estimating amplified band intensities. When NCS2 mutant callus lines cultured on regeneration medium, no adventitious shoot induction was observed. However, callus lines with more mitochondria induced adventitious shoots. These studies suggest that mitochondria NADH-dehydrogenase for electron transport in the inner membrane of mitochondria is essential for the differentiation and development of plants.

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Electrocatalytic Oxidation of NADH at the Modified Graphite Electrode Incorporating Gold Nano Particles (금 나노입자를 회합시킨 수식된 흑연전극으로 NADH의 전기촉매 산화반응)

  • Cha, Seong-Keuck;Han, Sung-Yub
    • Journal of the Korean Electrochemical Society
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    • v.10 no.1
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    • pp.1-6
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    • 2007
  • Mercaptopropionic acid(mpa) has been used to make self-assembled monolayer(SAMs) on the surface of graphite electrode incorporating gold nano particles, which are subsequently modified with dopamine(dopa). Such modified electrodes haying types of Gr(Au)/mpa-dopa were employed in the electrocatalytic oxidation of NADH. The responses of such modified electrodes were studied in terms of electron transfer kinetics and reaction procedure in the reaction. The reaction of the surface immobilized dopa with NADH was studied using the rotating disk electrode technique and a value of $5.06{\times}10^5M^{-1}s^{-1}$ was obtained for the second-order rate constant in 0.1 M phosphate buffer(pH=7.0), which was a $EC_{cat}$ and kinetic controlled procedure. But, the modified electrodes were diffusion controlled reaction having $4.64{\times}10^{-4}cm^2s^{-1}$ of the coefficient within $10^{-3}s$ after starting the reaction.

Metabolic Flux Shift of Weissella kimchii sk10 Grown Under Aerobic Conditions

  • Park, Sun-Mi;Kang, Hye-Sun;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.919-923
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    • 2004
  • The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of $NADH/NAD^+$, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.

Catalytic Oxidoreduction of Pyruvate/Lactate and Acetaldehyde/Ethanol Coupled to Electrochemical Oxidoreduction of $NAD^+$/NADH

  • Shin, In-Ho;Jeon, Sung-Jin;Park, Hyung-Soo;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.540-546
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    • 2004
  • We deviced a new graphite-Mn(II) electrode and found that the modified electrode with Mn(II) can catalyze NADH oxidation and $NAD^+$ reduction coupled to electricity production and consumption as oxidizing agent and reducing power, respectively. In fuel cell with graphite-Mn(II) anode and graphite-Fe(III) cathode, the electricity of 1.5 coulomb (A x s) was produced from NADH which was electrochemically reduced by the graphite-Mn(II) electrode. When the initial concentrations of pyruvate and acetaldehyde were adjusted to 40 mM and 200 mM, respectively, about 25 mM lactate and 35 mM ethanol were produced from 40 mM pyruvate and 200 mM acetaldehyde, respectively, by catalysis of ADH and LDH in the electrochemical reactor with $NAD^+$ as cofactor and electricity as reducing power. By using this new electrode with catalytic function, the bioelectrocatalysts are engineered; namely, oxidoreductase (e.g., lactate dehydrogenase) and $NAD^+$ can function for biotransformation without electron mediator and second oxidoreductase for $NAD^+$/NADH recycling.

Electrocatalytic Oxidation of NADH at Electrodes of Self-assembled Monolayers Modified with Dopamine (도파민으로 수식된 SAMs 전극에서 NADH의 전기촉매에 의한 산화)

  • Cha, Seong-Keuck
    • Journal of the Korean Chemical Society
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    • v.48 no.2
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    • pp.151-155
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    • 2004
  • The self-assembled mololayers(SAMs)were prepared with cysteine(cys) and subsequently coupled with dopamine(dopa) containing quinone functionality on the gold modified electrodes. The SAMs annealed in ethanol for 6 hours gave a better shaped cyclic voltammogram which had a 0.28 V of formal potential and same redox potential in 0.1M phosphate buffer(pH=7.10). The electrodes were employed to determine concentration of HADH with the result that calibration curve exhibited an excellent correlation(${\geq}$ 0.993) for the concentrations ranging up to 5.0${\times}10^{-4}$ M.

Biological activity of quinoline derivatives as inhibitors of NADH-ubiquinone oxidoreductase in the respiratory chain (NADH-ubiquinone oxidoreductase 저해제인 quinoline 유도체들의 생리활성)

  • Chung, Kun-Hoe;Cho, Kwang-Yun;Takahashi, Nobutaka;Yoshida, Shigeo
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.43-48
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    • 1991
  • New quinoline compounds were designed, synthesized, and examined with submitochondria. Most compounds showed high activity against NADH-ubiquinone oxidoreductase. Inhibition activity was mainly affected by the length of the lipophilic part, regardless of bulkiness or location of a phenyl group in the side chain. The $\beta-methyl$ group was demons)rated to be the optimal functionality on the nuclei of the quinoline derivatives so 4hat either deletion or insertion of a methylene on the group eliminated its activity.

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Simple Preparation of Diaphorase/Polysiloxane Viologen Polymer Modified Electrode for Sensing NAD and NADH

  • Song, Ji-Eun;Hong, Zhenyu;Nagarale, Rajaram Krishna;Shin, Woon-Sup
    • Journal of Electrochemical Science and Technology
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    • v.2 no.3
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    • pp.163-167
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    • 2011
  • Nicotinamide adenine dinucleotide, $NAD^+$, and its reduced form, NADH, play important roles as coenzymes in many enzymatic reactions. Electrochemical methods for $NAD^+$ or NADH detection or generation are drawn attention because it can provide the simple and low cost platform with fairly good sensitivity. In this study, the polysiloxane viologen polymer/diaphorase/hydrophilic polyurethane (PSV/DI/HPU) modified electrodes were simply prepared and demonstrated for bio-electrocatalytic $NAD^+$ sensors. The electrodes were co-immobilized with diaphorase and polysiloxane viologen polymer as an electron mediator followed by the overcoating with HPU membrane. The mixture of the enzyme and the electron mediator was well stabilized within HPU membrane and exhibited good reversibility and stability. The sensitivity was 0.2 $nA{\cdot}{\mu}M^{-1}$ and the detection limit was 28 ${\mu}M$ with a response time of 50 s ($t_{90%}$). The capability for NADH sensor was also observed on the PSV/DI/HPU electrode.

Covalent Immobilization of Diaphorase in Viologen Polymer Network for Highly Sensitive Detection of NAD+ and NADH

  • Song, Jieun;Hong, Zhenyu;Koh, Ahyeon;Shin, Woonsup
    • Journal of Electrochemical Science and Technology
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    • v.5 no.1
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    • pp.19-22
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    • 2014
  • Here we report a highly sensitive and stable detection of $NAD^+$ and NADH by the electrode on which diaphorase (DI) is covalently immobilized in viologen polymer network. The network is prepared by the covalent formation of the structure by mixing propylamine viologen (PAV), poly(ethylene glycol)(400) diglycidyl ether (PEGDGE), an diaphorase (DI). The PAV/PEGDGE/DI modified electrode has the sensitivity of $0.02nA{\cdot}{\mu}M$ and the detection limit of $3{\mu}M$ with a response time of 2 s ($t_{90%}$) for NADH sensing.

Type conversion of single walled carbon nanotube field effect transistor using stable n-type dopants

  • Yun, Jang-Yeol;Ha, Jeong-Suk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.268-268
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    • 2010
  • 단일벽 탄소 나노튜브(SWCNT)는 그 뛰어난 전기적, 물리적 특성 때문에 반도체 공정에 있어서 중요한 p-type 채널 물질로 꼽히고 있다. 본 연구에서는 SWCNT를 성장하여 이를 이용한 전계효과 트랜지스터를 제작하고 또한, 부분적인 폴리머의 코팅으로 타입을 변화하는 연구를 보이고자 한다. Ferritin용액을 DI-water에 2000배 희석하여 SiO2 기판 위에 뿌린 뒤 Methanol을 이용하여 기판 표면에 촉매가 붙어있게 한다. 이 기판을 $900^{\circ}C$로 가열하여 유기물질을 제거한 뒤 화학 기상 증착(Chemical Vapor Deposition)방법으로 SWCNT를 성장하게 된다. 이렇게 성장된 SWCNT는 촉매의 농도에 비례하는 밀도를 가지게 되며 이 위에 전극을 증착하고 back-gate를 설치하여 FET를 제작한다. 메탈릭한 SWCNT는 breakdown 공정을 통하여 제거한 뒤, 전자 농도가 높은 NADH를 전체적으로 코팅을 한다. NADH는 기존의 다른 폴리머(polyethyleneimine: PEI)에 비교하여 코팅 후 전자 제공 효과가 크며 그 성질의 재현성이 높고 공기 중에서 안정성을 유지하는 능력이 있다. 이러한 NADH의 코팅으로 n-type으로의 SWCNT FET를 제작하였으며 type conversion 현상을 이용하면 국부적인 NADH의 코팅으로 homojunction-diode의 제작 등 다양한 소자의 제작에 적용될 것으로 예상한다.

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