• Title, Summary, Keyword: glutamine

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Transformation of Glutamic Acid to Glutamine by E. coli Glutamine Synthetase (E. coli Glutamine Synthetase에 의한 Glutamic Acid로부터 Glutamine 으로의 전환)

  • 전복환;신형순;이왕식;방원기
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.379-384
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    • 1989
  • Glutamine production from glutamate was carried out using glutamine synthetase from E. coli K-12 pgln 6 and baker's yeast, which supplies ATP into the reaction system through alcohol fermentation, simultaneously. With whole cells of E. coli K-12 pgln 6 as an enzyme source of glutamine synthetase, 11.8 g/ι of glutamine produced after 18-h incubation (60% yield based on a substrate, glutamate). Using the partially purified glutamine synthetase, 19.8 git of glutamine was produced after 5-h incubation. This amount of glutamine was correspond to 90% yield, based on substrate, glutamate.

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Production of Glutamine by Glutamine Synthetase and Acetate Kinase of Escherichia coli (Escherichia coli의 Glutamine Synthetase와 Acetate Kinase에 의한 Glutamine 생산)

  • 조정일
    • The Korean Journal of Food And Nutrition
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    • v.6 no.3
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    • pp.169-177
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    • 1993
  • The conversion of glutamate by glutamine synthetase Is the endergonic reaction that demands ATP as its energy source. In order to supply efficiently ATP that is demanded in the conversion of glutamate to glutamine, the ATP- generating system by acetate kinase partially purified from Escherichia coli K-12 was coupled with glutamine synthetase partially purified 5. coli K-12 Pgln6. The optinum conditions of the coupled reaction were investigated. As the result, the highest conversion of glutamate to glutamine was shown In the reaction mixture containing 100mM glutamate, 100mM NHtCl, 50M acetyl phosphate, 5mM ADP, 40M MgCl2, 300mM potassium phosphate buffer (pH 7.5), 5mM MnCl2, Under this condition, the most effective concentrations of enzyme were 70unit/ml glutamine synthetase and 99unit/ml acetate kinase. Under the optinum conditions, 98% of 100mM glutamate was converted to glutamine within 6 hours.

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The Effect of Glutamine on Production of Tissue-type Plasminogen Activator from Recombinant Human Melanoma Cells in Glutamine-limited Fed-batch Cultivation

  • Kim, Hyun-Goo;Kim, Tae-Ho;Kim, Dae-Seok;Park, Kyung-Yu;Park, jin-seo;Ahn, Chol;Lee, Jin-Ha;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.295-298
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    • 1996
  • Under glutamine-limited condition, $2\times10^6$ (viable cells/ml) of maximum cell density and 13.5 ($\mu g$/ml) of tissue-type Plasminogen Activators (tPA) production were maintained by spike feeding fresh medium in fed-batch cultivation of human recombinant melanoma cells. It showed that tPA production was much seriously affected than cell growth according to initial glutamine concentrations. Above 3.4 (mmol/I) of glutamine concentration both cell growth and tPA production were not much affected by increasing initial glutamine concentration. Glutamine depleted situation was occurred at latter periods of batch and fed-batch cultivations below 5.4 (mmole/I) of initial glutamine concentration. It also showed that maximum glutamine consumption and ammonia evolution rates were closely related to initial glutamine concentrations. Maximum specific tPA production rate was estimated as $8.1\times19^{-6}$ ($\mu g$/cells/h) at 3.4(mmol/I) of glutamine concentration, which is higher than that from other batch and fed-batch processes.

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Light-dependent Hydrogen Production in Chlorobium limicola f. thiosulfatophilum NCIB 8327: A Possibility of Regulation via Glutamine Synthetase (Chlorobium limicola f. thiosulfatophilum NCIB 8327에서의 광수소발생 조절 기작에 대하여)

  • 나종욱;강사욱
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.558-563
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    • 1992
  • Chlorobium liimicola f. thiosulfatophilum NCIB 8327 was grown on modified Pfennig's medium using ammonium chloride. glutamine. glutamate, or dinitrogen gas as nitrogen sources. Except for the case of dinitrogen gas. the extent of gro\\1h was almost the s~me. The specific activity of glutamine synthetase in crude extracts is the highest in the cells which were grown on the medium containing glutamate. hut that of glutamate synthase is uniform for all four nitrogen sources. When the concentration of ammonium ions increases in the reaction mixture. the specific activity of glutamine synthetase in crude extract from the cells grown on glutamate decreases. hut that of glutamate dehydrogenase increases. whereas that of glutamate synthase remains unchanged. When the concentration of methionine sulfoximine increases, the activity of glutamine synthetases decreases rapidly. On the other hand. when the concentration of ammonium ions increases in the reaction mixture gradually. the activity of glutamine synthetase from the cells grown on higher concentration of ammonium ions less decreases. In the presence of light. the activity of glutamine synthetase increases. hut in the dark it decreases gradually. The production of hydrogen in intact cells depends on light. It is inhihited by adding ammonium ions. hut restores immediately hy adding methionine sulfoximine. The produclion of hydrogen in this strain can he mediated by nitrogenase only. and regulated hy glutamine synthetase.

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Effect of Glutamine on the Diclofenac Induced Bacterial Translocation and Lipid Peroxidation (Diclofenac에 의해 유발된 장내세균전위와 지질과산화에 대한 글루타민의 효과)

  • Kim, Eun-Jeong;Kim, Jeong-Wook
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.128-133
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    • 2005
  • The aim of this study was to examine whether administration of glutamine are able to prevent the NSAID induced bacterial translocation and lipid peroxidation in the rats. The an imals with glutamine were fed with L-glutamine for 5 days before diclofenac administration (100 mg/kg orally). 48 hour after diclofenac administration, intestinal permeability, serum biochemical profiles, and malondialdehyde levels of ileum were measured for evaluation of gut damage. Also, enteric aerobic bacterial counts, number of gram-negatives in mesenteric Iymph nodes, liver, spleen and kidney and malondialdehyde levels in liver, spleen, kidney and plasma were measured. Diclofenac caused the gut damage, enteric bacterial overgrowth, increased bacterial translocation and increased lipid peroxidation. Co-administration of glutamine reduced the gut damage, enteric bacterial overgrowth, bacterial translocation and lipid peroxidation induced by diclofenac. This study suggested that glutamine might effectively prevent non-steroidal anti-inflammatory drug induced bacterial translocation and lipid peroxidation in the rat.

Effects of Alanine and Glutamine Supplementation on Alcohol Metabolism in ICR Mice

  • Jonghee Chyun;Jungeun Yim;Youngnam Cha
    • Nutritional Sciences
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    • v.5 no.1
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    • pp.9-12
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    • 2002
  • This study was conducted to investigate the effects of oral supplementation of alanine and glutamine on alcohol metabolism. The subjects were 70 male ICR mice weighing 25-30 g. The animals were raised on standard rations artier weaning. After 24 hours of fasting, all the animals were given a peritoneal injection of 20% alcohol. Then, they were randomly divided into two groups: control and experimental. Fifteen minutes after the injection of alcohol, the mice in the experimental group wer given an oral solution of alanine(5 mM, 2 g/kg B. W) and glutamine (5 mM, 2g/kg B.W). The concentration of alcohol in the blood was measured in all the mice 20 minutes after they received the alochol, and the measurements continued every 20 minutes up to 140 minutes. The experimental group sustained lower blood alcohol levels at every 20 minute time interval compared to the control group, showing that oral supplementation of alanine and glutamine increases the rate of alcohol metabolism. Furthermore, the total amount of alcohol remaining in the blood, determined by using the Area Under the Curve (AUG) method, was lower in the group supplemented with alanine and glutamine, However, the effectiveness of alanine and glutamine in increasing the rate of alcohol metabolism, compared to the control group, diminished with time throughout the experiment. In conclusion, alanine and glutamine supplementation appears to promote alcohol metabolism shorthy after alcohol intake.

Studies on the Maturation of rabbit Follicular Oocytes in Vitro: Effects of Amino Acids and Carbohydrates

  • Bae, In-Ha
    • The Korean Journal of Zoology
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    • v.18 no.4
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    • pp.181-196
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    • 1975
  • Rabbit follicular oocytes were cultured in a basic medium containing 0.4% bovine serum albumin (BSA), carbohydrates and amino acids in various combinations. Osmolarity of the medium was maintained at 308 mOsm. The carbohydrates, pyruvate, lactate and glucose were all about equally beneficial, but not essential for rabbit oocyte maturation. Glutamine and proline, but not methionine or phenylalanine stimulated oocyte develoment. Glutamine stimulated more follicular oocytes to develop to prophase and metaphase II than did any of the three carbohydrates tested alone or in combination. Ammonia production after 24 hours of culture was highest in medium containing glutamine(15.2$\\mu$g/ml) but this was not inhibitory to maturation. Negligible amounts of ammonia were found with the other amino acids added. The optimum level of osmolarity for rabbit oocyte maturation appears to be ranged from 250$\\sim$310 mOsm with the maximum level of 270 mOsm. With 0, 0.08, 0.4, 2, 10 and 50 mM of glutamine in the medium, plus BSA but without carbohydrates, 30, 73, 70, 71, 59, 45% of the oocytes developed to prophase or metaphase II respectively. This indicates that no carbohydrate is required of the maturation of rabbit oocytes when 0.08$\\sim$2 mM of glutamine is included, which are the optimum range. Follicular oocytes could develop in the medium containing $^14 C$-glutamine and BSA but without carbohydrates or other organic compound. From the $^14 CO_2$ produced and TCA precipitable material isolated, it is suggested that glutamine probably is utilized by oocytes and cumulus cells as a source of energy as well as for protein synthesis.

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Effect of Methionine Sulfoximine in nitrogenase activity by ammonia and glutamine in Rhodopseudomonas sphaeroides (암모니아와 glutamine에 의한 Rhodopseudomonas sphaeroides의 질소 고정 효소활성에 미치는 Methionine Sulfoximine의 영향)

  • 이혜주
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.215-222
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    • 1988
  • The effect of ammonia and glutamine on nitrogenase activity of Rhodopseudomonas sphaeroides was examined. The nitrogenase activity of this strain was inhibited by ammonia and glutamine. When ammonia and glutamine were exhausted, nitrogenase activity promptly resumed at its original rate. Methionine sulfoximine (MSX), irreversible glutamine synthetase (GS) inhibitor, is a structural analogue of glutamate. MSX was used in order to know whether the nitrogenase activity was inhibited by ammonia and glutamine directly or not. The ability of MSX to prevent nitrogenase switch-off by ammonia was found to be dependent upon the phase of culture. When the cells were sampled after 12 hour culture, $500{\mu}M$ MSX would not prevent the nitrogenase switch-off by ammonia. Twenty one percents of GS actibity was inhibited by $500{\mu}M$ of MSX and concentration of released ammonia decreased. But nitrogenase activiy was still inhibited by ammonia. However, nitrogenase switch-off after 20 hours would be prevented by $100{\mu}M$ of MSX. On the other hand, GS activity was ingibited completely by $100{\mu}M$ MSX and concentration of released ammonia somewhat increased. But nitrogenase activity was not inhibited. The data indicated that the inhibition of in vivo nitrogenase actibity of Rp. sphaeroides by ammonia seemed to be mediated by products of ammonia assimilation rather than by ammonia itself.

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Glutamine Synthetase of some Fermentation Bacteria: Function and Application

  • Tachiki, Takashi
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • pp.506-508
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    • 1986
  • Metabolic activity of inorganic nitrogenous compounds affects not only microbial growth but also metabolite production in fermentation technology. We have worked on the enzymes participating in ammonia assimulation of some fermentation bacteria. This paper summarizes the results on glutamine synthetase and its application in practical field. Glutamine synthetase (L-glutamate:ammonia ligase, EC. 6.3.1.2) catalyzes the formation of glutamine from glutamate and ammonia at the expense of cleavage of ATP and inorganic phosphate. The enzyme plays a dual role in nitrogen metabolism in bacteria; it is a key enzyme not only in the biosynthesis of various compounds through glutamine but also in the regulation of synthesis of some enzymes involved in the metabolism of nitrogenous compounds. The detailed works with the Eschericia coli and other enterobacterial enzymes revealed that glutamine synthetase is controlled by the following complex of mechanisms: (a) feedback inhibition by end products, (b) repression and derepression of enzyme synthesis, (c) modulation of enzyme activity in response to divalent cation and (d) covalent modification of enzyme protein by adenylylation and its cascade control. Comparative studies have also been made on the enzymes from other organisms.

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Purification and Characterization of Extracellular and Intracellular Glutamine Synthetases from Mycobacterium bovis BCG

  • SUH, CHANG-IL;JUN-MAN LIM;HA-CHIN SUNG
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.946-950
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    • 2001
  • Slow-growing pathogenic mycobacterium species, including Mycobacterium bovis BCG, secrete a large amount of glutamine synthetase into culture media. Extracellular and intracellular glutamine synthetases were purified from M. bovis BCG. While the native molecular weights of both glutamine synthetases were estimated to be 370.2 kDa, those of the subunits were 61.7 kDa, indicating that the native forms were composed of 6 subunits. The enzymes showed a hhigh thermal stability and high degree of sequence similarity with the glutamine synthetase from M. tuberculosis in the N-terminal amino acid sequence. Western blotting analysis indicated that the antibodies prepared against both the extracellular and intracellular enzymes exhibited common antigen determinants.

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