• Title/Summary/Keyword: NAD(P)H

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Effects of Tributyltin in vitro on Hepatic Monooxygenase System in Marine Fishes (유기주석화합물이 해산 어류의 간장 MFO 효소계에 미치는 영향)

  • 전중균;이미희;이지선;심원준;이수형;허형택
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.18-25
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    • 2003
  • Effects of tributyltin chloride (TBTC) in vitro on mixed function oxygenase (MFO) system on liver microsome of eight marine fish species were investigated. To determine the effects on MFO system, cytochrome P45O (CYP) and cytochrome b5 con-tents, activities of two reductases (NADH-cytochrome b5 reductase and NADPH-cy-tochrome P450 reductase) and four dealkylation enzymes (EROD, PROD, MROD and ECOD) were measured in fish microsoms exposed to TBTC for 20 min. The WP content was reduced to 10% of the control group in 6 out of 8 species exposed to TBTC, whereas there was no significant change in the cytochrome bs content. the response of NAD(P)H dependant reductases depended on fish species. The dealkylation enzyme activities in microsome were also apparently inhibited by TBTC. The degree of inhibition was different among fish species and four enzymes. The EROD activities in eight species were decreased to the range of 1∼65% of control group.

Intracellular Concentrations of NAD(P), NAD(P)H, and ATP in a Simulated Oxic-settling-anaerobic (OSA) Process (OSA 공정의 세포 내 ATP, NAD(H), NADP(H) 농도)

  • Ventura, Jey-R Sabado;Nam, Ji-Hyun;Yang, Benqin;Na, Ri;Kil, Hyejin;Nam, Deok-Hyeon;Kang, Ki-Hoon;Jahng, Deokjin
    • Journal of Korean Society on Water Environment
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    • v.31 no.6
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    • pp.599-609
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    • 2015
  • In order to investigate why OSA (oxic-settling-anaerobic) process produces less sludge than CAS (conventional activated sludge) process, sequential cultivation through 1st aerobic-anaerobic-2nd aerobic conditions, were carried out. Then, the intracellular concentrations of adenosine triphosphate (ATP), nicotinamide adenine dinucleotide (NAD and NADH), and nicotinamide adenine dinucleotide phosphate (NADP and NADPH) were monitored for these three stages. Results showed that the concentrations of these energy substances rapidly decreased through time in both aerobic and anaerobic conditions but the anaerobic culture contained the lower energy level than aerobic culture. The 2nd aerobic culture that experienced anaerobic condition showed lower concentration of these energy substances than those of the 1st aerobic culture. Meanwhile, the anaerobic culture corresponding to the sludge holding stage of OSA was subjected to different soluble chemical oxygen demand (SCOD) levels, detention time, and temperature to evaluate the effects of these variations on the energy level difference between the 1st and 2nd aerobic stages. The lower the SCOD concentration, the longer detention time; and the higher temperature in the anaerobic stage tended to further reduce the intracellular level of the 2nd aerobic culture. On the average, the intracellular energy level of the anaerobic and 2nd aerobic stage were 57.73% and 39.12% of the 1st aerobic culture, respectively. These indicated that the insertion of an anaerobic stage between two aerobic stages could lower the intracellular energy levels, hence the lower the sludge in OSA than CAS process. Moreover, manipulation of the operating conditions of the intervening anaerobic stage can change intracellular energy levels thereby controlling sludge production.

Characterization of the Outer Membrane-Associated 2-Furaldehyde Dehydrogenase from Klebsiella pneumoniae (Klebsiella pneumoniae 균주의 세포외막에서 분리한 2-furaldehyde dehydrogenase의 특성에 관한 연구)

  • 이준우;강사욱;하영칠;한홍의
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.197-206
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    • 1988
  • An outer membrane-associated 2-furaldehyde dehydrogenase, catalyzing the oxidation of 2-furaldehyde to 2-furoic acid from Klebsiella pneumoniae was purified to homogeneity and characterized. The enzyme showed its highly specific dependency on $\beta$-$NAD^{+}$. Enzyme activity was monitored during purification by using substrate 2-furaldehyde and coenzyme $\beta$-$NAD^{+}$ by means of high performance liquid chromatography. The outer membrane was successfully collected by the methods of Percoll density gradient ultracentrifugation and ultracentrifugation after preferential solubilization of the membrane with $Mg^{2+}$ and Triton X-100. The enzyme was purified by the series of procedures including extraction of outer membrane protein with EDTA and lysozume, and fractionation by column chromatography on QAE-Sephades Q-50, and subsequently Sephadex G-100. The enzume showed its optimal activity at $85^{\circ}C$, pH 9.5, and in the presence of 1.5% (vol/vol) Triton X-100. The enzyme exhibited a native molecular size of 88,000 by nondenaturing polyacrylamide gel electrophoresis and had an apparent Km of 4.72mM for 2-furaldehyde.

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Arabidopsis SIZ1 positively regulates alternative respiratory bypass pathways

  • Park, Bong-Soo;Kim, Sung-Il;Song, Jong-Tae;Seo, Hak-Soo
    • BMB Reports
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    • v.45 no.6
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    • pp.342-347
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    • 2012
  • Plant mitochondria possess alternative respiratory pathways mediated by the type II NAD(P)H dehydrogenases and alternative oxidases. Here, E3 SUMO ligase was shown to regulate alternative respiratory pathways and to participate in the maintenance of carbon and nitrogen balance in Arabidopsis. The transcript abundance of the type II NAD(P)H dehydrogenases NDA2 and NDB2 and alternative oxidases AOX1a and AOX1d genes was low in siz1-2 mutants compared to that in wild-type. The addition of nitrate or ammonium resulted in a decrease or an increase in the expression of the same gene families, respectively, in both wild-type and siz1-2 mutants. The amount of free sugar (glucose, fructose and sucrose) was lower in siz1-2 mutants than that in wild-type. These results indicate that low nitrate reductase activity due to the AtSIZ1 mutation is correlated with an overall decrease in alternative respiration and with a low carbohydrate content to maintain the carbon to nitrogen ratio in siz1-2 mutants.

Induction of NAD(P)H:quinone reductase and glutathione S-transferase by Xanthii Fructus and Prunellae Spica Extracts (창이자 및 꿀풀하고초에 의한 NAD(P)H:quinone reductase와 glutathione S-transferase의 유도)

  • Shon, Yun-Hee;Lee, Ki-Taek;Park, Sin-Hwa;Cho, Kyoung-Hee;Lim, Jong-Kook;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.32 no.4 s.127
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    • pp.269-273
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    • 2001
  • Ethanol extracts from Xanthii Fructus (XFE) and Prunellae Spica (PSE) were investigated for the effects on the induction of cancer chemoprevention-associated enzymes. The following effects were measured: (a) induction of quinone reductase (QR) (b) induction of glutathione S-transferase (GST) (c) reduced glutathione (GSH) level. XFE and PSE were potent inducers of quinone reductase activity in Hepa1c1c7 murine hepatoma cells. Glutathione levels were increased with XFE and PSE. In addition, glutathione S-transferase activity was increased with XFE. However, GST activity was not increased with PSE. These results suggest that XFE and PSE have chemopreventive potentials by inducing quinone reductase and increasing GSH levels.

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Effects of Xylose Reductase Activity on Xylitol Production in Two-Substrate Fermentation of Recombinant Saccharomyces cerevisiae

  • Lee, Woo-Jong;Kim, Myoung-Dong;Yoo, Myung-Sang;Ryu, Yeon-Woo;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.725-730
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    • 2003
  • Three recombinant Saccharomyces cerevisiae strains showing different levels of xylose reductase activity were constructed to investigate the effects of xylose reductase activity and glucose feed rate on xylitol production. Conversion of xylose to xylitol is catalyzed by xylose reductase of Pichia stipitis with cofactor NAD(P)H. A two-substrate fermentation strategy has been employed where glucose is used as an energy source for NADPH regeneration and xylose as substrate for xylitol production. All recombinant S. cerevisiae strains Yielded similar specific xylitol productivity, indicating that xylitol production in the recombinant S. cerevisiae was more profoundly affected by the glucose supply and concomitant It generation of cofactor than the xylose reductase activity itself. It was confirmed in a continuous culture that the elevation of the glucose feeding level in the xylose-conversion period enhanced the xylitol productivity in the recombinant S. cerevisiae.

Two-enzyme coupled fluorometric assay of urinary dipeptidase (이원효소 연쇄반응의 형광분석에 의한 Urinary Dipeptidase의 활성도 측정)

  • Park, Haeng Soon;We, Jeoung Soon
    • Analytical Science and Technology
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    • v.8 no.3
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    • pp.359-364
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    • 1995
  • Urinary dipeptidase(Udpase) was assayed by fluorometric analysis of NADH which was produced from an indicator enzyme, L-alanine dehydrogenase. The reaction mixture was consisted of a dipeptide(L-ala-L-ala), ${\beta}-NAD^+$, L-alanine dehydrogenase in 12.5 mM sodium carbonate buffer, pH 9.0, and urinary dipeptidase which initiated the reaction. The fluorescence intensity of NADH was measured as a function of time with the excitation wavelength at 340nm and emission at 460nm. Comparison of this fluorometric method with the conventional spectrophotometric method utilizing glycyldehydrophenylalanine(Gdp) as substrate provided the correlation coefficient of 0.996 and increased the sensitivity more than ten times.

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Induction of Glutathione S-transferase and NAD(P)H:Quinone Reductase by Astragali Radix Aqua-acupuncture Solution (황기(黃耆) 약침액(藥鍼液)의 Glutathione S-transferase 와 NAD(P)H: Quinone Reductase 유도)

  • Ryu Jun-Seon;Lim Jong-Kook
    • Korean Journal of Acupuncture
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    • v.18 no.1
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    • pp.21-26
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    • 2001
  • Induction of phase II enzymes such as quinone reductase (QR) or glutathione S-transferase (GST) is considered a major mechanism of protection against initiation of carcingenesis. This study was desinged to investigate the potential of Astragali Radix Aqua-acupuncture Solution (ARAS) to induce phase II enzymes and glutathione (GSH) in murine hepatoma cells grown in microtiter plate wells. ARAS was potent inducers of QR activity. ARAS was induced about 2.6-fold at concentration of $5{\times}$. In addition, GST activity was increased with ARAS. GSH levels were increased about 1.2-fold with ARAS at concentration of $0.1{\times}$. These results suggested that ARAS may act as blocking agents against carcinogenesis by induction of phase II marker enzymes.

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Iron Toxicity to Peritoneal Macrophage Due to Alteration of Mitochondria by NO

  • Yoon, Ji-Yeon;Kim, Jin-Sun;Lee, Heum-Sook;Lee, Kyo-Young;Cheon, Choong-Ill;Lee, Myeong-Sok;Park, Jong-Hoon;Song, Eun-Sook
    • Animal cells and systems
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    • v.8 no.2
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    • pp.97-103
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    • 2004
  • The cytotoxic effect of iron was examined in peritoneal macrophage to determine contributing factors by iron injection to rat. Viability was reduced by 24% by the iron-overload and by 30% by short-term iron addition. Total iron was increased by 45% in the iron-overloaded with remarkable elevation (9 to 14 fold) in the presence of $FeSO_4$. Free calcium was also increased by 19% in control and 44% in iron-overloaded group due to additional $FeSO_4$ NO and MDA were increased by 40% and 136%, respectively, with significant reduction (37%) of NAD(P)H. RCR and cytochrome c oxidase activity were lowered approximately by 10% with reduction of mitochondrial membrane potential. Addition of iron was frequently associated with altered distribution of mitochondria of high membrane potential in the iron-overloaded macrophage. These results suggest altered mitochondria with high NO and low NAD(P)H due to iron.