• Title/Summary/Keyword: oxaloacetate

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Low Serum Medium for CH07E02 Hybridoma (하이브리도마 배양을 위한 저혈청배지의 조성 결정)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.473-477
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    • 1993
  • Up to now, 10% Fetal Bovine Serum(FBS(V/V)) was added to basal medium for the cultivation of hybridoma. For the cultivation of hybridoma cell line, CH07E02, against colon cancer, serum concentration was reduced to 3% FBS without influence on cell growth and maximum cell concentration. By the addition of cell growth promoting substances-insulin (I), pyruvate (P), oxaloacetate(O), Pluronic F-68(P) and 2-mercaptoethanol(2-ME)-to 1% FBS medium, a cell density higher than that with 1% FBS medium alone was achieved. FBS 3% medium was replaced by very cheap 2% Calf Serum (CS) medium without influence on cell growth rate and concentration. Cells grew vigorously in 0.5% CS+IPOP medium. This composition was used during suspension culture and exhibited good viability and high specific growth rate.

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Effect of 6-Aminonicotinamide on the Levels of Some Metabolites and Related Enzymes in Rabbit Serum (6-Aminonicotinamide가 토끼혈청내 효소 및 대사물질에 미치는 영향)

  • Park, In-Koo;Lee, Chul-Seung;Lee, Seung-Hoon;Song, Yoon-Kyung;Shin, Sook
    • The Korean Journal of Zoology
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    • v.33 no.4
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    • pp.493-498
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    • 1990
  • The effects of an antimetabolite, 6-aminonicotinamide (6-AN) on the levels of enzymes and metabolites in rabbit serum were investigated. The intraperitoneal administration of 6-AN (multiple doses of l5mg/kg body weight) gave tise to a remarkable increase in glucose and cholesterol levels but did not exert any appreciable influence on the concentration of albumin and total protein. Alkaline phosphatase activity was significantly reduced by administration of 6-AN, whereas creatine phophokinase, serum glutamic oxaloacetate transaminase and serum glutamic pyruvate transaminase activities were matkedly enhanced. Nevettheless, the levels of Ca, P, Na, K, Cl and Co were not affeded to any extent by 6-AN.

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Deregulation of Aspartokinase by Single Nucleotide Exchange Leads to Global Flux Rearrangement in the Central Metabolism of Corynebacterium glutamicum

  • Kim Hyung-Min;Heinzle Elmar;Wittmann Christoph
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1174-1179
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    • 2006
  • The wild-type Corynebacterium glutamicum ATIC 13032 and Corynebacterium glutamicum ATTC 13032 lysC S301Y, exhibiting a deregulated aspartokinase, were compared concerning growth, lysine production, and intracellular carbon fluxes. Both strains differ by only one single nucleotide over the whole genome. In comparison to the wild-type, the mutant showed significant production of lysine with a molar yield of 0.087 mol (mol glucose$^{-1}$) whereas the biomass yield was reduced. The deregulation of aspartokinase further led to a global rearrangement of carbon flux throughout the whole central metabolism. This involved an increased flux through the pentose phosphate pathway (PPP) and an increased flux through anaplerosis. Because of this, the mutant revealed an enhanced supply of NADPH and oxaloacetate required for lysine biosynthesis. Additionally, the lumped flux through phosphoenolpyruvate carboxykinase and malic enzyme, withdrawing oxaloacetate back to the glycolysis and therefore detrimental for lysine production, was increased. The reason for this might be a contribution of malic enzyme to NADPH supply in the mutant in the mutant. The observed complex changes are remarkable, because they are due to the minimum genetic modification possible, the exchange of only one single nucleotide.

Chemical Modification of Residue of Lysine, Tryptophan, and Cysteine in Spinach Glycolate Oxidase

  • Lee, Duk-Gun;Cho, Nam-Jeong;Choi, Jung-Do
    • BMB Reports
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    • v.29 no.4
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    • pp.321-326
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    • 1996
  • Spinach glycolate oxidase was subjected to a series of chemical modifications aimed at identifying amino acid residues essential for catalytic activity. The oxidase was reversibly inactivated by treatment with pyridoxal 5'-phosphate (PLP). The inactivation by PLP was accompanied by the appearance of an absorption peak of around 430 nm, which was shifted to 325 nm upon reduction with $NaBH_4$. After reduction, the PLP-treated oxidase showed a fluorescence spectrum with a maximum of around 395 nm by exciting at 325 nm. The substrate-competitive inhibitors oxalate and oxaloacetate provided protection against inactivation of the oxidase by PLP. These results suggest that PLP inactivates the enzyme by fonning a Schiff base with lysyl residue(s) at an active site of the oxidase. The enzyme was also inactivated by tryptophan-specific reagent N-bromosuccinimide (NBS). However, competitive inhibitors oxalate and oxaloacetate could not protect the oxidase significantly against inactivation of the enzyme by NBS. The results implicate that the inactivation of the oxidase by NBS is not directly related to modification of the tryptophanyl residue at an active site of the enzyme. Treatments of the oxidase with cysteine-specific reagents iodoacetate, silver nitrate, and 5,5'-dithiobis-2-nitrobenzoic acid did not affect significantly the activity of the enzyme.

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Effects of Nicotinic Acid Deficiency on the Levels of Various Metabolites in the Serum of Quail (Nicotinic acid 결핍이 메추리 혈청의 여러 대사물질 수준에 미치는 효과)

  • 이재혁;박인국
    • The Korean Journal of Zoology
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    • v.34 no.2
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    • pp.203-208
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    • 1991
  • Body weight gain in the niadn deficient group of quail was markedly lowered as compared to that of the control group, but heart, kidney and liver weight were slighdy reduced reladve to the body weight. Nicotinic acid deficiency resulted in the significant increase of serum glucose level but the serum cholesterol, albumin and total protein levels were not affected to any extent. Glutamic oxaloacetate iransaminase and glutamic pyruvate transaminase activities were significandy enhanced but alkaline phosphatase and lactic dehydrogenase activities were not influenced. Tryptophan and tyrosine levels were remarkably reduced and a similar observation was also made with aspartic acid, glutamic acid and alanine plus serine. However, the levels of basic amino acids such as arginine, histidine and lysine plus branched chain amino acids such as isoleucine, leucine and valine were not affected.

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Aspartate-glutamate carrier 2 (citrin): a role in glucose and amino acid metabolism in the liver

  • Milan Holecek
    • BMB Reports
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    • v.56 no.7
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    • pp.385-391
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    • 2023
  • Aspartate-glutamate carrier 2 (AGC2, citrin) is a mitochondrial carrier expressed in the liver that transports aspartate from mitochondria into the cytosol in exchange for glutamate. The AGC2 is the main component of the malate-aspartate shuttle (MAS) that ensures indirect transport of NADH produced in the cytosol during glycolysis, lactate oxidation to pyruvate, and ethanol oxidation to acetaldehyde into mitochondria. Through MAS, AGC2 is necessary to maintain intracellular redox balance, mitochondrial respiration, and ATP synthesis. Through elevated cytosolic Ca2+ level, the AGC2 is stimulated by catecholamines and glucagon during starvation, exercise, and muscle wasting disorders. In these conditions, AGC2 increases aspartate input to the urea cycle, where aspartate is a source of one of two nitrogen atoms in the urea molecule (the other is ammonia), and a substrate for the synthesis of fumarate that is gradually converted to oxaloacetate, the starting substrate for gluconeogenesis. Furthermore, aspartate is a substrate for the synthesis of asparagine, nucleotides, and proteins. It is concluded that AGC2 plays a fundamental role in the compartmentalization of aspartate and glutamate metabolism and linkage of the reactions of MAS, glycolysis, gluconeogenesis, amino acid catabolism, urea cycle, protein synthesis, and cell proliferation. Targeting of AGC genes may represent a new therapeutic strategy to fight cancer.

Analysis of Productivity of a Polyculture Container for Suspended Rearing of Abalones and Sea cucumbers (전복과 해삼 수하양식을 위한 복합사육기의 생산력 분석)

  • Nam, Myung-Mo;Lee, Chu;Moon, Tae-Seok;Kim, Su-Kyung;Hwang, Jin-Wook
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.323-330
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    • 2011
  • We have developed a polyculture container which is preferable for rearing of abalones and sea cucumbers in East Sea. To test the rearing capacity of the polyculture container, 50, 75 and 100 sea cucumbers ranging from the body length of 7-9 cm were cultured in three containers including 500 abalones with the shell length of about 5 cm, respectively. It was revealed that preferable density for sea cucumbers was 52-72 individuals in the polyculture container. Glutamate oxaloacetate transaminase and glutamate pyruvate transaminase were analyzed with Reflotron kit to investigate the health degree of abalones in two polyculture containers including 300 and 500 abalones with the shell length of about 5 cm. Glutamate oxaloacetate transaminase has been an important tool to know myocardial infarction, disease of liver, and destruction of muscle. Next experiment was conducted to determine the effects on growth of abalones and sea cucumbers in polyculture container. Experimental findings, RNA/DHA ratio, DNA and RNA contents (ug/mg) were not significantly different among all groups. The results imply that the method of polyculture can be rearing with sea cucumber without growth retardation of abalone. Production ability between polyculture container and the container used in south sea were carried out using suspended culture method during 8months. It was revealed that abalones and sea cucumbers are faster growing in polyculture container than in container used in south sea. Therefore, polyculture container is considered more appropriate for the abalones and sea cucumbers culture in East Sea.

Studies on the Origin of Prunus yedoensis Matsumura 1. A comparative Electrophoretic Study on Wild P. subhirtella in Mt. Hanla, Cultivated P. yedoensis and P. donarium (왕벚의 기원에 관한 연구 1. 한라산자생올벚, 재배왕벚 및 산벚의 전기영동형 비교)

  • 한창열
    • Journal of Plant Biology
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    • v.20 no.1
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    • pp.1-5
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    • 1977
  • Horizontal starch gel electrophoresis in three buffers was used to compare the electrophoretic patterns in three cherry species, wild Prunus subhirtella, cultivated P. yedoensis and P. donarium. Electrophoretic patterns of glutamate oxaloacetate, transaminase-2(GOT-2), malate dehydrogenase-2(MDH-2), and phosphoglucose isomerase(PGI) in three species showed strong evidence that P. yedoensis might be originated by hybridization between P. subhirtella and P. donarium.

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Protective Role of Light in Heat-Induced Inhibition of Photosynthesis in Isolated Chloroplasts

  • Jun, Sung-Soo;Kim, Chang-Hoon;Hong, Young-Nam
    • Journal of Photoscience
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    • v.5 no.4
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    • pp.157-162
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    • 1998
  • The effect of heat treatment in the light on the subsequent CO2 fixation was studied with isolated spinach chloroplasts to define the role of light during heat stress. The degree of inhibition in CO2 fixation after heat treatment at 35$^{\circ}C$ under full light intensity (600W/$m^2$) was same as that in the dark. However, heat treatment of isolated chloroplasts in the light manifested thylakoidal damage, which did not occur in the dark. Under weak light (10~30 W/$m^2$ ) where no thylakoidal damage occurred, the inhibition was substantially alleviated , showing protective effect of light . The inhibition caused by heat treatment in the dark or light is prevented by the addition of a few combined compounds to the medium prior to treatment. Fructose-1-6- bisphosphate(with aldolase)and ribose-5-phosphate, known to be effective combined with oxaloacetate in preventing inhibition after heat treatment in the dark were equally effective in the light even without oxaloacetate. Addition of sugar phosphate reduced the Mehler reaction, which may occur in fast rae under high light. However, the addition of bicarbnate and catalase that would remove Mehler reaction did not provide any protection, indicating that protective role of sugar phosphate is elsewhere. Furghermore, in whole plants rapid recovery from heat stress was observed in the light. The apparently lesser or equal inhibition in spite of additional thylakoidal damage under heat stres in the light and less requirement for the protection against heat treatment suggest that the inhibitory effect of heat stress is alleviated by light treatment.

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Formation of Succinic Acid by Klebsiella pneumoniae MCM B-325 Under Aerobic and Anaerobic Conditions

  • Thakker Chandresh;Bhosale Suresh;Ranade Dilip
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.870-879
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    • 2006
  • The present study describes the formation of succinic acid by a nonvirulent, highly osmotolerant Klebsiella pneumoniae strain SAP (succinic acid producer), its profile of metabolites, and enzymes of the succinate production pathway. The strain produced succinate along with other metabolites such as lactate, acetate, and ethanol under aerobic as well as anaerobic growth conditions. The yield of succinate was higher in the presence of $MgCO_3$ under $N_2$ atmosphere as compared with that under $CO_2$ atmosphere. Analysis of intracellular metabolites showed the presence of a smaller PEP pool than that of pyruvate. Oxaloacetate, citrate, and $\alpha$-ketoglutarate pools were considerably larger than those of isocitrate and fumarate. In order to understand the synthesis of succinate, the enzymes involved in end-product formation were studied. Levels of phosphoenolpyruvate carboxykinase, fumarate reductase, pyruvate kinase, and acetate kinase were higher under anaerobic growth conditions. Based on the profiles of the metabolites and enzymes, it was concluded that the synthesis of succinate took place via oxaloacetate, malate, and fumarate in the strain under anaerobic growth conditions. The strain SAP showed potential for the bioconversion of fumarate to succinate under $N_2$ atmosphere in the presence of $MgCO_3$. At an initial fumarate concentration of 10 g/l, 7.1 g/l fumarate was converted to 7 g/l succinate with a molar conversion efficiency of 97.3%. The conversion efficiency and succinate yield were increased in the presence of glucose. Cells grown on fumarate contained an 18-fold higher fumarate reductase activity as compared with the activity obtained when grown on glucose.