• Title/Summary/Keyword: ATP synthesis efficiency

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Efficiency of ATP Synthesis and Impairment of Glucose Tolerance in the NIDDM-Prone Rat

  • Kim, Sook-Bae
    • Journal of Nutrition and Health
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    • v.30 no.4
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    • pp.379-385
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    • 1997
  • This study was designed to determine whether genetic defects in the efficiency of ATP synthesis existed in the NIDDM-prone BHE/cdb rat and to determine whether these defects caused the development of glucose intolerance. Thyroxine treatment provided an excellent clue as to the nature of the genetic defects in this rat. The characteristics of hyperhyroid and control Sprague-Dawley(SD) and BHE/cdb rats were studied. Hyperthyroidism was induced through the addition of thyroxine($T_4$) to the diet(2mg/kg of diet). Active proton conductances and passive proton conductances were tested. Mitochondria from hyperhyroid BHE/cdb rats were less efficient iii active proton conductances than mitochondria from hyperhyroid SD rats. It showed that decreased efficiency of ATP synthesis in the BHE/cdb rat was probably related to defects in active proton conductance, Indicating aberrant FoATPase. The levels of $F_1F_0$ATPaseATPase activity were tested. Mitochondria from hyperthyroid BHE/cdb rats were less active than mitochondria from hyperthyroid SD rats. This may be an attribute of aberrant F$_1$ATPase and may contribute to the BHE/cdb strain s characteristic of reduced ATP synthesis efficiency. Glucose tolerances were tested. BHE/cdb rats were profoundly affected by thyroxine, whereas SD rats were less so. It showed that the diabetes phenotype in BHE/cdb rats was related to defects in thyroxine-induced uncoupling. These results showed the decreased efficiency of ATP synthesis due to genetic defects in $F_1F_0$ATPase had relevance to the characteristic of impaired glucose tolerance in the NIDDM-prone BHE/cdb rat.

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Enhancement of Glucose-Fueled Cell-Free Protein Synthesis by the Addition of Lipids (지질의 첨가를 통한 포도당 기반 무세포 단백질 합성 시스템의 단백질 발현 효율 향상)

  • Lee, So Jeong;Kim, Ho-Cheol;Kim, Dong-Myung
    • Korean Chemical Engineering Research
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    • v.57 no.1
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    • pp.85-89
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    • 2019
  • Cell-free protein synthesis utilizes the translational machinery in a cell extract. Unlike the conventional cell-based expression methods, not being affected by the conditions for cell growth, cell-free protein synthesis enables flexible manipulation of individual factors affecting the efficiency protein biosynthesis. However, the high cost and low stability of the energy sources to regenerate ATP have limited the use of cell-free synthesis for large-scale production of recombinant proteins. One of the approaches to address this problem is to use glucose as an alternative energy source to regenerate ATP through the glucose-metabolizing pathways in a cell extract. In this study, in an attempt to improve the efficiency of ATP regeneration by reinforcing oxidative phosphorylation process, we supplemented with cellular lipids to a glucose-fueled reaction mixture for cell-free protein synthesis. As a result of the lipid supplementation, the productivity of chloramphenicol acetyltransferase in a cell-free synthesis system using glucose increased more than 6 fold compared to when the lipid was not supplemented.

Metabolic Flux Analysis of a Poly-${\beta}$-hydroxybutyrate Producing Cyanobacterium, Synechococcus sp. MA19, Grown under Photoautotrophic Conditions

  • Nishioka, Motomu;Nishiuma, Hajime;Miyake, Masato;Asada, Yasuo;Shimizu, Kazuyuki;Taya, Masahito
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.295-302
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    • 2002
  • To understand the utilization property of light energy, Synechococcus sp. MA19, a poly-${\beta}$-hydroxybutyrate (PHB) producer, was cultivated at the different incident light intensities of 15.3, 50.0 and 78.2 W/$m^2$ using media with and without phosphate. From the results of metabolic flux analysis, it was found that the cell yield based on ATP synthesis was estimated as $3.5{\times}10^{-3}$ kg-biomass/mol-ATP in these cultures. Under the examined conditions, there were no significant differences in the efficiency of light energy conversion to chemical energies estimated as ATP synthesis and reducing potential (NADH + NADPH) formation whether the PHB synthesis took place or not. The energy converted from light to ATP was kept relatively high around the energy absorbed by the cells of $2.5-3.0{\times}10^{6} J\;h^{-1}\;kg^{-1}$, whereas the energy of reducing potential was hardly changed in the examined range of the energy absorbed by the cells.

Regeneration of ATP through an Activated Glycolytic Pathway in a Cell-free Extract and its Application for Protein Expression (해당과정의 활성화를 통한 무세포 단백질 발현 시스템에서의 ATP 재생)

  • Kim Dong Myung;Keum Jeong Won;Kim Tae Wan;Oh In Seok;Choi Cha-yong
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.467-470
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    • 2004
  • We have investigated the key parameters affecting ATP regeneration in a cell-free protein synthesis system derived from Escherichia coli. When glucose-6-phosphate was used as an energy source, the efficiency of ATP regeneration sharply responded to pH change of reaction mixture. In addition, both productivity and reproducibility of protein synthesis was substantially enhanced by introducing appropriate amount of NAD into the reaction mixture. As a result, through the activation of glycolytic pathway under an optimal pH, the batch cell-free system produced over $300\;{\mu}g$ of protein in a 1 mL reaction.

Effects of Taurine Supplementation on Mitochondrial Function in Chronic Ethanol Administered Rats

  • Shim Kwan-Seop;Park Garng-Hee;Kim Sook-Bae
    • Journal of Community Nutrition
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    • v.7 no.3
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    • pp.163-168
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    • 2005
  • The present investigation was undertaken in vivo to determine whether the functional alterations of hepatic mitochondria induced by ethanol might be prevented by taurine. We examined the effects of supplementation of taurine on hepatic mitochondrial oxidative phosphorylation in the chronic ethanol-administered rats. Isolated hepatic mitochondria from three groups of rats were functionally tested by an analysis of $\beta-hydroxbutyrate-supported$ respiration and the coupling of this process to ATP synthesis in the presence of ADP. The three groups were control group(CO), ethanol(60g/L) administered group (AL), and ethanol (60g/L) + taurine (5g/L) supplemented group (AT). Ethanol and/or taurine were given in drinking water for 10 weeks. The mitochondria from AL group had lower state 4 respiratory rate, respiratory control (RC) ratio and ADP : O(P/O) ratio than those from CO and AT group. It showed that the ethanol administered rats were less coupled and thus less efficient with respect to mitochondrial ATP synthesis than both control rats and ethanol + taurine supplemented rats. It suggests that taurine supplementation might improve the impaired oxidative phosphorylation efficiency in mitochondrial dysfunction that is recognized as a cause of liver diseases in chronic ethanol consumption.

Effects of Exercise and Supplementation of L-Carnitine and Antioxidants on Mitochondrial Function in Rats

  • Kim, Sook-Bae;Kim, Sun-Ju
    • Journal of Community Nutrition
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    • v.4 no.3
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    • pp.187-194
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    • 2002
  • This study was investigated the effects of exercise and supplementation of L-carnitine and antioxidants on hepatic mitochondrial function, especially oxidative phosphorylation (OXPHOS). Isolated hepatic mitochondria from 4 rat groups were functionally tested by an analysis of respiration and the coupling of this process to ATP synthesis in the presence of ADP. Four groups were non-trained, non-supplemented group (NTNS), non-trained, supplemented group (NTS), long term-trained, non-supplemented group (LTNS) , and long term-trained, supplemented group (LTS). The trained rats run on a treadmill (grade 10°,20 m/min) for 60min/day for 8 weeks. The supplemented rats were treated with L-carnitine (0.5% diet), vitamin E(0.5mg/g BW), vitamin C (0.5mg/g BW) and melatonin (1 $\mu$ g/g BW) for 8 weeks. There were exercise effects on improving mitochondrial OXPHOS. Within non-supplemented groups, exercised rats resulted in a significant decrease in state 4 oxygen consumption, which increased the respiratory control (RC) ratio and ADP : O (P/O) ratio. There were supplementation effects on improving mitochondrial OXPHOS, too. Within non-exercised rats, supplemented rats resulted in a significant decrease in state 4 oxygon consumption. which increased the RC ratio and P/O ratio. There were additive effects of exercise and supplementation on OXPHOS. Within supplemented rats, exercise resulted in an increase in RC ratio. Significant effects of exercise-supplement interaction on improving OXPHOS were identified. It suggests that exercise and supplementation of L-carnitine and antioxidants might improve more efficiently the impaired OXPHOS efficiency in mitochondrial dysfunction that recognized as is an important cause of degenerative diseases. (J Community Nutrition 4(3) : 187∼194, 2002)