• Title/Summary/Keyword: pyruvate accumulation

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Induction of antioxygenic enzymes as defense systems in plant cells against low temperature stress : (I) Accumulation of pyruvate in cells during cold treatment and activation of antioxygenic enzymes during post-chilling period (식물의 냉해에 대한 생체방어기구로서 항산소성 효소의 유도 : (1) 저온처리중 pyruvate의 세포내 축적과 상온환원후 항산소성 효소의 활성화)

  • Kim, Jong-Pyung;Hahn, Chang-Kyun;Jung, Jin
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.162-167
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    • 1991
  • In an attempt to explore the mechanistic aspects of chilling injury in plants and their defensive measures against the low temperature stress, the time sequential measurements of pyruvate, superoxide radicals$(O_{\overline{2}})$ and antioxygenic enzymes during whole period of injury-inducing treatment were performed using mostly rice seedlings. Pyruvate was substantialy accumulated in leaf tissues during the exposure period to $5^{\circ}C$ of the seedlings ; the relative extent of the accumulation was increased with increasing time of the cold treatment. When the cold-treated plants were translocated to ambient temperature$({\sim}25^{\circ}C)$, the accumulation started to dissipate, concomitantly accompaning a remarkable increase in the $O_{\overline{2}}$ level of tissues. Superoxide dismutase(SOD) and catalase were also activated during post-chilling period, although they showed a considerable lag time for activation. In contrast, glutathione peroxidase, another antioxygenic enzyme in cells, was not activated at all by preceding cold treatment of plants. The uptake of exogenous $O_{\overline{2}}$ by the roots of rice seedlings resulted in increase in the activities of SOD and catalase in root tissues. The supply of $H_2O_2$ to plan st brought about the activation of catalase in situ, while failing to exert any effect on the activation state of glutathione peroxidase. The results obtained in this work suggest that pyruvate accumulation in cells is the direct cause of the overproduction of $O_{\overline{2}}$ and thereby other toxic activated oxygen species, and that SOD and catalase may play a crucial role in the protection of plant cells against active oxygen-mediated chilling injury.

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A Novel Therapeutic Measure for Metabolic Acidosis with Amino Acids

  • Kim, Jun;Goo, Yong-Sook;Kim, Sang-Jeong;Park, Sang-Chul;Koh, Chang-Soon
    • The Korean Journal of Physiology
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    • v.26 no.1
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    • pp.89-97
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    • 1992
  • In hypoxic tissue conditions, pyruvate can not enter the Krebs cycle and lactic acid, produced from pyruvate, accumulates to induce lactic acidosis. Pyruvate, However, can also be converted to alanine by glutamate pyruvate transaminase, that could be enhanced by glutamate. Therefore, it would be a fundamental measure to treat the lactic acidosis in tissue hypoxic conditions when one can convert the accumulated lactic acid, through pyruvate, to alanine. To test the above hypothesis, we induced a lactic acidosis in cats and the effect of glutamate on recovery of acid base state and removal of the lactic acid from blood were assessed and the results were compared with those of bicarbonate administration, which is one of the most frequently used conventional measure for correction of the acid base state during lactic acidosis. The results were that glutamate and combined glutamate bicarbonate solutions not only restored the acid base status completely from the lactic acidosis in an hour or two, but also restored the blood level of lactate partially. We concluded that administration of glutamate solution to convert pyruvate into alanine is effective in preventing lactic acid accumulation and treating lactic acidosis.

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A Mechanistic Study on the Early Stage-Events Involved in Low Temperature Stress in Clamydomonas reinhardtii (Clamydomonas reinhardtii의 냉해 초기과정에 관한 기작론적 연구)

  • Cho, Hyun-Soon;Kim, Chang-Sook;Jung, Jin
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.433-440
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    • 1994
  • The exposure of Clamydomonas reinhardtii to low temperatures resulted in an accumulation of cellular pyruvate that dissipated when the chilled cells returned to ambient temperature. The dissipation of pyruvate accumulation was accompanied by an increase in the production level of superoxide radicals $(O_2^-)$ in cells. The formation of $O_2^-$ at an excessive level during the post-chilling period was apparently countered by a substantial activation of superoxide dismutase (SOD). All these results are similar to those observed previously in rice seedlings subjected to the cold-treatment, implicating that a common mechanism is probably underlying for the primary processes of chilling injury both in higher plants and in algae. It was also observed that the activation of Mn-containing SOD contributes the major share in the increase of SOD activity of whole algal cells. Because Mn-SOD is present only in mitochondria, the observation corroborates the concept that the $O_2^-$ scavenging enzyme would be induced to cope with the cold treatment-caused adverse situation in mitochondria where the toxic active oxygen is produced at rates far exceeding the normal rate.

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Analysis of the Growth and Metabolites of a Pyruvate Dehydrogenase Complex-Deficient Klebsiella pneumoniae Mutant in a Glycerol-Based Medium

  • Xu, Danfeng;Jia, Zongxiao;Zhang, Lijuan;Fu, Shuilin;Gong, Heng
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.753-761
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    • 2020
  • To determine the role of pyruvate dehydrogenase complex (PDHC) in Klebsiella pneumoniae, the growth and metabolism of PDHC-deficient mutant in glycerol-based medium were analyzed and compared with those of other strains. Under aerobic conditions, the PDHC activity was fourfold higher than that of pyruvate formate lyase (PFL), and blocking of PDHC caused severe growth defect and pyruvate accumulation, indicating that the carbon flux through pyruvate to acetyl coenzyme A mainly depended on PDHC. Under anaerobic conditions, although the PDHC activity was only 50% of that of PFL, blocking of PDHC resulted in more growth defect than blocking of PFL. Subsequently, combined with the requirement of CO2 and intracellular redox status, it was presumed that the critical role of PDHC was to provide NADH for the anaerobic growth of K. pneumoniae. This presumption was confirmed in the PDHC-deficient mutant by further blocking one of the formate dehydrogenases, FdnGHI. Besides, based on our data, it can also be suggested that an improvement in the carbon flux in the PFL-deficient mutant could be an effective strategy to construct high-yielding 1,3-propanediol-producing K. pneumoniae strain.

Effect of Chemical Properties of Cultivation Soils on the Plant Growth and the Quality of Garlic (재배지 토양의 화학성이 마늘의 생육 및 품질에 미치는 영향)

  • Kim, Chang-Bae;Kim, Chan-Yong;Park, Man;Lee, Dong-Hoon;Choi, Jyung
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.5
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    • pp.333-339
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    • 2000
  • Effects of chemical properties of cultivation soils on the growth and quality of garlics were investigated. Garlics were cultivated in Uisung and Yechun, one of the major areas of garlic production, where upland and paddy fields have been used for garlic production for many years. Contents of phosphate, sulfur and potassium in the soils of paddy fields were relatively higher than those in the soils of upland fields, suggesting that the accumulation of inorganic salts has been progressed in the paddy fields. Soils of Uisung area showed higher pH s and lower contents of available phosphate compared to those of Yechon area. This result implies that the soils of Uisung area provide somewhat better chemical properties for garlic growth than those of Yechun area. Contents of inorganic salts such as phosphate, potassium and magnesium in the soils significantly affected the growth and quality of garlics. Garlics grown in the soils with lower contents of these inorganic salts exhibited better growth status and contained more pyruvate. More pyruvate was found in the garlics grown in upland fields than in paddy fields. Therefore, it is apparent that the accumulation of inorganic salts, especially available phosphate, in cultivation soils leads to the inhibition of garlic growth and in turn to the deterioration of garlic quality.

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Growth Characteristics of a Pyruvate Decarboxylase Mutant Strain of Zymomonas mobilis (Pyruvate decarboxylase 돌연변이 Zymomonas mobilis 균주의 생장 특성 연구)

  • Xun, Zhao;Peter L., Rogers;Kwon, Eilhann E.;Jeong, Sang Chul;Jeon, Young Jae
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1290-1297
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    • 2015
  • Studies of the inactivation of a gene encoding pyruvate decarboxylase, pdc, in an ethanol-producing bacterium, Zymomonas mobilis, identified a mutant strain with 50% reduced PDC activity. To evaluate the possibility of a carbon-flux shift from an ethanol pathway toward higher value fermentation products, including pyruvate, succinate, and lactate, fermentation studies were carried out. Despite attempts to silence pdc expression in the wild-type strain ZM4 using cat-inserted pdc and pdc-deleted homologs by electroporation, the strain isolated showed partial gene activation. Fermentation experiments with the PDC mutant strain showed that the reduced expression level of PDC activity resulted in decreased rates of substrate uptake and ethanol production, together with increased pyruvate accumulation of 2.5 g l-1 , although lactate and succinate concentrations were not significantly enhanced in these modified strains. Despite numerous attempts, no strains were isolated in which complete pdc inactivation occurred. This result indicates that the ethanol fermentation pathway of this bacterium is totally dependent on the activity of the PDC enzyme. To ensure a redox balance of intracellular NAD and NADH levels, other enzymes, such as lactate dehydrogenase for lactate, and enzymes involved in the production of succinic acid, such as pyruvate dehydrogenase (PDH) and malic enzymes, may be needed for their increased end-product production.

Determination of Optimal Toxic Concentration and Accumulation of Cadmium in Broiler Chicks

  • Subhan, Fazli;Khan, Ayaz;Wahid, Fazli;Shehzad, Adeeb;Jan, Amin Ullah
    • Toxicological Research
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    • v.27 no.3
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    • pp.143-147
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    • 2011
  • Cadmium is considered one of the most toxic, non biodegradable heavy metal for the human and animals. The purpose of the present study was to investigate the changes in biochemical parameters of blood and accumulation of cadmium in various tissue caused by various levels of dietary cadmium chloride ($CdCl_2$) in broiler chicks. $CdCl_2$ was administered through drinking water to broiler chicks. In spectral analysis, $CdCl_2$ treatment caused a significant increase in Glutamate pyruvate transaminase (GPT), creatinine and uric acid levels in all treated groups. Intriguingly, the GPT, creatinine, and uric acid levels were significantly higher at 75 mg/kg as compared to the groups treated with high doses (100, 125 and 150 mg/kg) of $CdCl_2$. Atomic Absorption Spectrophotometer (AAS) was used for the determination of Cd accumulation in kidney, liver and Breast muscles. AAS analysis revealed that Cd accumulation is increased in breast muscles as compared to liver and kidney at higher doses of Cd than 75 mg/kg.

Effects of Glucose and Acetic Acid on the Growth of Recombinant E.coli and the Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody (유전자 재조합 대장균의 세포성장과 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체 생산에 대한 포도당과 초산의 영향)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.482-488
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    • 2000
  • The Fab fraction of PDC-E2 specific human monoclonal antibody was produced using recombinant E. coli, and the effects of glucose and acetate were investigated to develop an optimal strategy for recombinant human antibody production. Higher glucose concentration in the culture media resulted inn higher cell growth and glucose consumption rate, which in turn resulted in an increased acetate production rate. When glucose was depleted, cells began to consume acetate as an energy source, and this consumption rate depended on the glucose concentration. When the residual glucose concentration was high, the accumulation of acetate was accelerated due to an increase in the acetate production rate and a decrease in the acetate consumption rate. Futhermore, it was found that a high accumulation of acetate, accompanied by a high glucose concentration, inhibited human antibody formation; the critical acetate concentration was $0.6g/\ell$. During production, a high glucose concentration enhanced cell growth, but inhibited antibody formation due to catabolic repression. Therefore, it is important to keep the concentration of both glucose and acetate as low as possible to increase antibody production after induction. Accordingly, it is important to accurately control the concentration of glucose and acetate in the culture media to obtain high cell densities and high productivity levels of recombinant human antibody.

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Metabolism of $C^{14}-1-glucose$ and $C^{14}-6-glucose$ by the Ehrlich Ascites Turner Tissue (에르릿히 복수종양의 $C^{14}-1-$ 포도당 및 $C^{14}-6-$포도당 대사에 관한 연구)

  • Kwon, Chang-Rak
    • The Korean Journal of Physiology
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    • v.1 no.1
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    • pp.33-41
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    • 1967
  • The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.

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Effect of Dipeptides on In vitro Maturation, Fertilization and Subsequent Embryonic Development of Porcine Oocytes

  • Tareq, K.M.A.;Akter, Quzi Sharmin;Tsujii, Hirotada;Khandoker, M.A.M. Yahia;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.501-508
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    • 2013
  • The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode's albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2-4 cell, 8-16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.