• Title/Summary/Keyword: glutathione synthetase

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Studies on the Properties of E. coli ${\gamma}-Glutamylcysteine$ Synthetase in Relation to the Enzymatic Synthesis of Glutathione (글루타치온의 효소적 생합성에 관계되는 E.coli ${\gamma}-Glutamylcysteine$ Synthetase의 특성 연구)

  • Nam, Yong-Suk;Kwak, Joon-Hyeok;Lee, Se-Yong
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.478-483
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    • 1997
  • ${\gamma}-Glutamylcysteine$ synthetase was purified from E. coli K-12 strain and its properties related to the in vitro synthesis of glutathione by enzymatic method were investigated. The activity of purified ${\gamma}-glutamylcysteine$ synthetase was increased with increasing concentration of L-glutamate up to 60 mM, while it was decreased by about 50% and 40% under 60 mM of L-cysteine and 45 mM of glycine, respectively. The enzyme activity was reduced not only by ADP, one of the reaction products, but also by the reduced form of glutathione. Therefore, because the reduced glutathione as well as glycine which is the substrate for glutathione synthetase inhibit the activity of ${\gamma}-glutamylcysteine$ synthetase, it is recommended to design a bioreactor system with two separate reactions for glutathione synthesis : one with ${\gamma}-glutamylcysteine$ synthetase reaction and the other glutathione synthetase reaction. In addition since ADP, resulted from these reactions, reduces the activity of ${\gamma}-glutamylcysteine$ synthetase, it is necessary to introduce an ATP regeneration system for glutathione synthesis.

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백서에서 Paraquat가 장기 Glutathione 농도에 미치는 영향

  • Go, Chun Nam;Go, Chun Nam;Yu, Ui Gyeong
    • Journal of the Korean Chemical Society
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    • v.38 no.1
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    • pp.69-73
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    • 1994
  • The effects of paraquat administration on glutathione was studied in rats. The contents of glutathione in the liver, kidney and lung were significantly decreased but the alteration was not significant in blood by paraquat administration. The decrease occured without concomitant increases in oxidized glutathione (GSSG) or in the GSH/GSSG ratio. The activities of γ-glutamylcysteine synthetase in liver and kidney were decreased by paraquat administration. And γ-glutamyl transpeptidase activities were significantly decreased in kidney and lung of paraquat treated-rats. These results suggest that the decreased synthesis of glutathione by paraquat were an important mechanism of the decreased level of glutathione in liver and kidney, and decreased glutathione transport was a factor on the changes of glutathione contents in lung.

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Effects of Bromate on the Glutathione Synthesis in Various Organs of Rats (Bromate가 흰쥐의 장기 Glutathione 함량에 미치는 영향)

  • 김나영;강혜옥;이무강;최종원
    • Journal of Life Science
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    • v.13 no.5
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    • pp.626-633
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    • 2003
  • The effects of bromate administration on glutathione were studied in rats. The contents of glutathione in the liver and kidney were significantly decreased but the alteration was not significant in lung and blood by bromate adminstration. The decrease occurred without concomitant increases in oxidized glutathione (GSSG) or in the GSSG/GSH+GSSG ratio. The activities of $\gamma-glutamyl$ cysteine synthetase in the liver and kidney were decreased by bromate administration. $\gamma-Glutamyl$ transpeptidase activities was significantly decreased in the kidney and not significantly decreased in the lung of bromate treated-rats. These results suggest that the decreased synthesis of glutathione by bromate may be an important reason for the decreased level of glutathione in the liver and kidney, thus the decreased glutathione transport would be a factor on the changes of glutathione contents in bromate-treated rats.

Glutathione Content and the Activities of Glutathione-Synthesizing Enzymes in Fission Yeast are Modulated by Oxidative Stress

  • Lee, Yuk-Young;Kim, Su-Jung;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.248-251
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    • 2003
  • Glutathione (GSH) is an important factor in determining tolerance against oxidative stress in living organisms. It is synthesized in two sequential reactions catalyzed by ${\gamma}$-glutamylcysteine synthetase (GCS) and glutathione synthetase (GS) in the presence of ATP. In this work, the effects of three different oxidative stresses were examined on GSH content and GSH-related enzyme activities in the fission yeast Schizosaccharomyces pombe. GSH content in S. pombe was significantly enhanced by treatment with hydrogen peroxide, ${\beta}$-naphthoflavone (BNF) and tert-butylhydroquinone (BHQ). Simultaneously, they greatly induced GCS and GS activity. However, they did not have any effects on glutathione reductase activity. These results suggest that GCS and GS activities in S. pombe are up-regulated by oxidative stress.

In-vitro Production of Glutathione Using Yeast ATP Regeneration System and Recombinant Synthetic Enzymes from Escherichia coli. (효모의 ATP 재생산계와 대장균 유래의 재조합 생산효소를 이용한 in vitro 글루타치온 생산)

  • 고성영;구윤모
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.213-220
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    • 1998
  • An ATP regeneration system was used for the production of glutathione which was synthesized by a sequential action of ${\gamma}$-glutamyl-cysteine synthetase and glutathione synthetase. The synthetases above were produced in the recombinant E. coli (TG1/pDG7) with the highest specific production yield of 31 mg glutathione/g wet cell. Bakers yeast was considered to have economically a better ATP regeneration system although the glutathione production yield was lower than that of acetate kinase. It was also observed that the ATP regeneration system of bakers yeast was superior to that of Saccharomyces cerevisiae ATCC24858. The yield of glutathione production with bakers yeast was 36% with the ATP concentration of 5 mM. To avoid the cysteine limitation during the early phase of glutatione production, an extra cysteine was added at 2 hours after reaction and the production yield increased 1.91 times. The effectiveness of bakers yeast as an ATP regeneration system was proved by several sets of extra feeding experiments. The product inhibition by glutathione above 14 mM was also observed.

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Effects of Constituent Amino Acids of Glutathione and Ammonium Sulfate added to Hydroponic Solution on the Synthesis of Glutathione in Lettuce

  • Kim Ju-Sung;Seo Sang-Gyu;Kim Sun-Hyung;Usui Kenji;Shim Le-Sung
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.195-202
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    • 2005
  • The effects of constituent amino acids of glutathione (GSH), glutamate (Glu), cysteine (Cys) and glycine (Gly), on GSH synthesis in lettuce seedlings were examined in this study. The GSH concentration of the seedlings was increased to 5.1-fold and 1.6-fold the concentration of the control in the first leaves and roots, respectively, by simultaneous application of these constituent amino acids (Glu+Cys+Gly) at 100 mg/l to the culture solution for two days. In the first leaves and roots of these seedlings, the concentration of GSH was 180.4 and 14.6 nmole/gFW, and non-essential amino acids including Glu, Cys and Gly occupied 93.2% and 84.0% of the total free amino acids, respectively. Application of Cys greatly increased the concentration of GSH in the roots, and application of 50 mg/l Cys increased it to 26.1-fold the concentration in the control. The activity of GSH synthetase was higher in the leaves than in the roots, whereas the activity of ${\gamma}$-glutamylcysteine synthetase was higher in the roots than in the leaves.

The Changes of Antioxidant Enzymes in the Lung of Alloxan-induced Diabetic Rats (알록산 유도 당뇨흰쥐의 폐에서 황산화계의 변화)

  • 최형호;고광삼;임동윤
    • YAKHAK HOEJI
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    • v.39 no.6
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    • pp.654-660
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    • 1995
  • The present study was attempted to investigate the mechanism of oxidative cellular injuries which occur in diabetic rats by determining changes of antioxidant enzymes activity in the lung of alloxan-induced diabetic rats, the contents of glutathione in the lung, liver, blood samples, and ${\gamma}$-glutamylcysteine synthetase activities in the liver. Superoxide dismutase activities (SOD), including Cu, Zn-SOD and Mn-SOD, decreased in the lung of diabetic rats compared with those of normal control rats. However, activities of catalase and glutathione peroxidase(GPX) activities were not affected in the lung of diabetic rats. In diabetic rats, glutathione contents in the lung, liver, and blood samples, as well as the activities of ${\gamma}$-glutamylcysteine synthetase in the livers which is known to be the key enzyme of glutatione biosynthesis, decreased significantly. From these experimental results, it is thought that the decrease in SOD activities in the lung, glutathione contents and ${\gamma}$-glutamylcysteine synthetase activities in some tissues in alloxan-induced diabetic rats may be the crucial cause of vullnerability to oxidative cellular injuries.

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Construction Various Recombiant Plasmids for the Enhancement of Glutathione Production in E. coli. (E. coli에서 글루타치온 생산 증가를 위한 재조합 플라스미드의 구성)

  • 남용석;이세영
    • Journal of Life Science
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    • v.7 no.4
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    • pp.253-261
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    • 1997
  • In order to enhance glutathione production, various recombinant plasmids containing gshI and/or gshII genes isolated from E. coli K-12 were constructed and introduced into E. coli. Some plasmids contained one to three copies of gshI genes in pBR325 and others contained both gshI and genes for glutathione biosynthesis. $\gamma$-Glutamylcysteine synthetase activities of E, coli strains amplified tandem repeated gshI genes were dependent on the number of inserted gshI genes. The glutathione productivity of E. coli strains harboring various plasmids was investigated using an E. coli acetate kinase reaction as an ATP regenerating system. The glutathione productivity of E. coli strains harboring tandem repeated gshI genes was increased in proportion to the number of inserted gshI genes. By the introduction of gshII gene, the glutathione productivity of the E. coli was increased by two-fold compared with E. coli strain amplified gshI gene only. The enzymatic production of glytathione in E. coli was mainly affected by the increase of $\gamma$-glutamylcysteine synthetase activity. The highest glutathione productivity was obtained in E. coli strains harboring pGH-501 plasmid containing two copies of gshI and copy of gshII genes in pUC8 vector.

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Regulation of the Gene Encoding Glutathione Synthetase from the Fission Yeast

  • Kim, Su-Jung;Shin, Youn-Hee;Kim, Kyung-Hoon;Park, Eun-Hee;Sa, Jae-Hoon;Lim, Chang-Jin
    • BMB Reports
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    • v.36 no.3
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    • pp.326-331
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    • 2003
  • The fission yeast cells that contained the cloned glutathione synthetase (GS) gene showed 1.4-fold higher glutathione (GSB) content and 1.9-fold higher GS activity than the cells without the cloned GS gene. Interestingly, $\gamma$-glutamylcysteine synthetase activity increased 2.1-fold in the S. pombe cells that contained the cloned GS gene. The S. pombe cells that harbored the multi copy-number plasmid pRGS49 (containing the cloned GS gene) showed a higher level of survival on solid media with cadmium chloride (1 mM) or mercuric chloride ($10\;{\mu}M$) than the cells that harbored the YEp357R vector. The 506 bp upstream sequence from the translational initiation point and N-terminal8 amino acid-coding region were fused into the promoteriess $\beta$-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pUGS39. Synthesis of $\beta$-galactosidase from the fusion plasmid pUGS39 was significantly enhanced by cadmium chloride and NO-generating S-nitroso-N-acetylpenicillamine (SNAP) and sodium nitroprusside (SN). It was also induced by L-buthionine-(S,R)-sulfoximine, a specific inhibitor of $\gamma$-glutamylcysteine synthetase (GCS). We also found that the expression of the S. pombe GS gene is regulated by the Atf1-Spc1-Wis1 signal pathway.