• 한국응용곤충학회지
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• 제34권3호
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• pp.181-190
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• 1995

암컷 Aedes aegypti의 난성숙과장에서 새로 나타나는 malate dehydrogenase(L-malate, $NAD^+$ oxidoreductase, EC 1.11.37, MDH)를 DEAE-Sepharose, Sulphonyl-Sepharose, Cibacron 3FGA affinity chromatography를 이용하여 분리정제하여 그 특성을 조사하였다. 분자량은 70,000 dalton정도의 dimer 형태로 되어 있으며 최적 pH는 malate-oxaloacetate반응에서는 pH 9.0~9.2, oxaloacetate-malate 반응에서는 pH 9.8~10.2이었다. 정재된 MDH는 mitochondria에 위치하고 있으며 기질로서 malate에 대한 Km값의 경우 $1.29 \times 10^{-4}$ M, oxaloacetate에 대한 Km 값은 $6.58\times 10^{-4}$M, NAD에 대한 Km값은 $0.76\times 10^{-3}$ M이며 NADH에 대한 Km 값은 $3.8\times 10^{-3}$ M 을 보이고 있으며 각각의 기질에 의한 저해현상을 보이고 있었다. 기질에 대한 Km값을 부분적으로 분리한 DEAE-sepharose에 흡착된 원형질 MDH와 비교한 결과 malate에 대한 Km 이 $8.92\times 10^{-3}$으로 상당한 차이를 보이고 있었다. 또한 정제된 MDH는 cltrate, $\alpha$-ketoglutarate, ATP 등의 대사산물에 의하여 저해작용을 받았다. ATP 및 citrate에 의한 MDH 활성도 저해는 oxaloacetate-malate반응에서 보다는 malate-oxaloacetate 반응에서 덜 일어났다. Oxaloacetate-malate 반응의 경우 ATP에 의하여저해작용이 완전히 일어났으며 malate-oxaloacetate반응에서는 cltrate에 의하여 저해작용이 일어나지 않았다. 흡혈 후 생성되는 MDH는 난소에서 합성되며 흡혈 수 난소에서 18시간 때부터 활성도가 나타나 48시간 이후 최고 활성도가 유지되는데 TCA회로의 isocitrate dehydrogenase 의 경우 난소내에서의 활성도 변화가 MDH의 변화 양상과 같았다.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권2호
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• pp.91-95
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• 2009

The presence of considerable amount of enzymes of TCA cycle isocitrate dehydrogenase (ICDH-NADP+, EC1.1.1.42), $\alpha$-ketogluterate dehydrogenase ($\alpha$-KGD, EC1.2.4.2) and malate dehydrogenase (MDH, EC1.1.1.37) in fresh control and in vitro starved adult Isoparorchis hypselobagri establish the functional TCA cycle in this fluke. The major metabolic end products are pyruvate, lactate, oxaloacetate and malate. The ratio of oxaloacetate/malate assess that oxaloacetate is reduced to malate and in this fluke the reverse TCA cycle is active. The pyruvate/lactate ratio shows pyruvate is reduced to lactate and the fluke is homolactate farmenters.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권2호
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• pp.136-141
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• 2005

The capability of utilizing a TCA cycle intermediates as the sole carbon source by the multi-vitamin auxotrophic yeast Torulopsis glabrata CCTCC M202019 was demonstrated with plate count method. It is indicated that T. glabrata could grew on a medium with one of the TCA cycle intermediates as the sole carbon source, but more colonies were observed when glucose, acetate and one of the TCA cycle intermediates coexisted in the medium. Among the intermediates of the TCA cycle examined in this study, cell growth was improved by supplementing oxaloacetate. Further investigation showed that the presence of acetate was necessary when oxaloacetate was supplemented. By supplementing with 10 g/L of oxaloacetate in pyruvate batch fermentation, dry cell weight increased from 11.8 g/L to 13.6 g/L, and pyruvate productivity was enhanced from $0.96\;gL^{-1}h^{-1}\;to\;1.19 gL^{-1}h^{-1}$ after cultivation of 56 h. The yield of pyruvate to glucose was also improved from 0.63 g/g to 0.66 g/g. These results indicate that under vitamins limitation, the productivity and yield of pyruvate could be enhanced via an increase of cell growth by the supplementation of oxaloacetate.

• 한국미생물·생명공학회지
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• 제34권1호
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• pp.35-39
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• 2006

높은 C4 대사활성을 보이는 반추위미생물이 가지는 포도당 발효대사 조절양식의 한가지를 대장균에서 모사하였다. 대장균은 glycolytic condition에서는 phosphoenolpyruvate(PEP) ${\leftrightarrow}$ oxaloacetate(OAA)간 반응을 phosphenolpyruvate carboxylase(PPC)에 의해, gluconeogenetic condition에서는 phosphoenolpyruvate carboxykinase(PCK)에 의해 촉매하도록 조절한다. 반면 반추위미생물은 glycolytic condition에서 PCK를 통하여 반응이 촉매된다. 이러한 조절양식의 차이점이 C4 대사활성에 미치는 영향을 조사하기 위하며 ppc가 돌연변이되고 대신 인위적으로 PCK를 발현할 수 있는 대장균을 제조하였다. 이렇게 PEP-OAA간 대사조절이 변이된 대장균 K12 ppc-/pck+는 야생형 K12보다 2.5배의 높은 C4대사활성을 보였다. 대장균에서의 C4 대사생리를 증가시키는 연구는 대사공학을 이용한 여러가지 유용물질(i.e. 숙신산, ALA)생산에 응용하기 위한 기초자료로 활용될 수 있을 것으로 기대된다.

• 미생물학회지
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• 제25권3호
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• pp.189-197
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• 1987

배지성분으로 첨가된 glutamate의 농도가 균의성장고 tylosin 생합성에 미치는 영향을 조사하였다. 그 결과 oxaloacetate를 공동기질로 사용하는 효소의 활성에 의하여 균의 성장과 tylosin의 생합성이 조절됨을 알았다. 즉, citrate synthase와 aspartate aminotransferase의 활성은 균의 성장에 아주 긴요하며 methylmalonyl-CoA carboxytransferase의 활성은 tylosin 생합성에 아주 중요한 효소임을 알았다. Glutamate의 농도는 우의 효소의 활성에 직접적으로 영향을 주고 있음을 알았다.

• 한국식품영양과학회지
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• 제29권2호
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• pp.193-197
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• 2000

The effects of black soybena extracts on enzymes activies of rat were evaluated in present study. Eighty-four male Sprague-Dawley rats weighing 100$\pm$10g were divided into twelve groups which consisted of black soybean extract, Pb and Cd solution, and black soybean extract plus Pb or Cd soln groups. The weight gain was increased in black soybean extracts and Pb soln solution group but decreased in Cd soln solution group. The results obtained form the experiment were as follows: Glutamate pyruvate trasaminase (GPT) and glutamate oxaloacetate oxaloacetate transaminase (GOT) activities were not significantly different among experimental groups. The lactate dehydrogenase (LDH) activities of black soybean extract administered groups were decreased than those of Pb and Cd solution group. Black soybean group increased cholinesterase (ChEase) activity as compared to administration of Pb and Cd soln group.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.287-292
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• 1997

The role of glyoxylate bypass in lysine production by Corynebacterium glutamicum ssp. lactofermentum ATCC21799 was analyzed by using cloned aceA and aceB genes which encode enzymes catalyzing the bypass. Introduction of a plasmid carrying aceA and aceB to the strain increased enzyme activities of the bypass to approximately 5 fold on acetate minimal medium. The strain with amplified glyoxylate bypass excreted 25% more lysine to the growth medium than the parental strain, apparently due to the increased availability of intracellular oxaloacetate. The final cell yield was lower in the strain with amplified glyoxylate bypass. These changes were specific to the lysine-producing C. glutamicum ssp. lactofermentum ATCC21799, since the lysine-nonproducing wild type Corynebacterium glutamicum strain grew faster and achieved higher cell yield when the glyoxylate bypass was amplified. These findings suggest that the lysine producing C. glutamicum ssp. lactofermentum ATCC21799 has the ability to efficiently channel oxaloacetate, the TCA cycle intermediate, to the lysine biosynthesis pathway whereas lysine-nonproducing strains do not. Our results show that amplification of the glyoxylate bypass efficiently increases the intracellular oxaloacetate in lysine producing Corynebacterium species and thus results in increased lysine production.

• 대한한의학회지
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• 제23권2호
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• pp.139-150
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• 2002

OBJECTIVES : Schizandra chinensis is well known for its efficacy at liver function reinforcement, relieving thirst and recovery from fatigue. In this study, we examined the effects of sports drink containing Schizandra chinensis on serum metabolic substrate, electrolyte, stress indicators, related-enzyme and exercise performance, rectal temperature, and heat shock proteinb70 (HSP70). METHODS : Elite long-distance runners (male, 21.3yrs, n=16) were selected and divided into two groups; an experimental group (EXP, n=8) and a control group (CON, n=8). A beverage containing Schizandra chinensis was supplemented 3 times per day to EXP for 4 weeks. Serum biochemical elements (glucose, lactate, total cholesterol, triglyceride, high density lipoprotein cholesterol, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, creatinine, creatine phosphokinase, lactate dehydrogenase, blood urea nitrogen, Na, K, Cl) were analyzed by auto blood analyzer. Exercise performance was measured by treadmill exercise test, HSP70 was detected by electrophoresis and Western blotting, and rectal temperature was measured by rectal temperature probe. RESULTS : Administration of the beverage increased significantly the rest level of blood Na, Cl and glucose and decreased significantly lactate dehydrogenase, glutamate oxaloacetate transaminase. No difference was found in exercise performance, rectal temperature increment or HSP70 concentration between groups. CONCLUSIONS : Administration of a sports drink containing Schizandra chinensis altered blood glucose, lactate dehydrogenase, glutamate oxaloacetate transaminase, Na and Cl levels.

• Fisheries and Aquatic Sciences
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• 제5권4호
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• pp.308-310
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• 2002

Effect of 4-nonylphenol (4-NP), endocrine disrupting compounds (EDCs), on glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were investigated in the plasma of juvenile rockfish, Sebastes schlegeli. Fish were injected with 4­NP (10, 50, 100 and 200 mg/kg body weight) in $70\%$ ethanol twice at 3-day intervals and plasma sampling were extracted at 7 days after the last injection. Controls received solvent only. 4-NP significant increased GOT in a dose-dependent manner. GPT was markedly elevated to $61\%$ (P<0.05) and $82\%$ (P<0.01) than that of the control at the 4-NP doses of 100 and 200mg, respectively. These results suggest that the estrogenic activity of 4-NP increase plasma GOT and GPT by toxic effect on hepatocyte.

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.315-320
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• 1996

The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.