• Title/Summary/Keyword: Glutathione synthetase

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Transcriptional Regulation of the Gene Encoding ${\gamma}$-Glutamylcysteine Synthetase from the Fission Yeast Schizosaccharomyces pombe

  • Kim, Su-Jung;Kim, Hong-Gyum;Kim, Byung-Chul;Kim, Kyunghoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.233-238
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    • 2004
  • Transcriptional regulation of the Schizosaccharomyces pombe y-glutamylcysteine synthetase (GCS) gene was examined using the two GCS-lacZ fusion plasmids pUGCS101 and pUGCS102, which harbor 607 bp and 447 bp upstream regions, respectively. The negatively-acting sequence was located in the -607 - -447 bp upstream region of the GCS gene. The upstream sequence responsible for induction by menadione(MD) and L-buthionine-(S, R)-sulfoximine (BSO) resides in the -607 - -447 bp region, whereas the sequence which codes for nitric oxide induction is located within the -447 bp region, measured from the translational initiation point. Carbon source-dependent regulation of the GCS gene appeared to be dependent on the nucleotide sequence within -447 bp region. The transcription factor Papl is involved in the induction of the GCS gene by MD and BSO, but not by nitric oxide. Induction of the GCS gene occurring due to low glucose concentration does not depend on the presence of Pap1. These data imply that induction by MD and BSO may be mediated by the Pap1 binding site, probably located in the -607 - -447 region, and also that the nitric oxide-mediated regulation of the S. pombe GCS gene may share a similar mechanism with its carbon-dependent induction.

Cloning of Genes for the Biosynthesis of Glutathione from E. coIi K-12 (E.coli K-12 균주로부터 글루타치온 합성 유전자의 클로닝)

  • 남용석;박영인;이세영
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.575-582
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    • 1991
  • To increase the production of glutathione by the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were isolated and cloned. To clone a gshI gene, the GS903 mutant strain, which is deficient in $\gamma$-glutamylcysteine synthetase activity, has been raised. A gshI gene was cloned using pBR322 plasmid as a 3.6 Kb PstI DNA fragment isolated from E. coli K-12 chromosomal DNA. Also a gshIl gene was cloned using pUC13 plasmid as a 2.2 Kb PstI-BamHI DNA fragment. To study the effects of plasmid copy number and passenger DNA size on the expression levels of the gsh genes, various recombinant plasmids containing different sets of genes were constructed. The expression levels of the gsh genes were increased approximately twice higher in pUC series plasmids than that in pBR322 plasmid. But the sizes of the passenger DNA containing the gsh genes in the vector plasmid did not affect on the expression levels of the gsh genes.

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A Study on the Effects of Sungshimsan on the Heart Lipid Peroxide and Metabolic Enzyme System in Senescence Induced Rats (노화과정(老化過程)의 흰쥐에서 성심산(醒心散)이 심장(心臟)의 대사효소계(代謝酵素系)에 미치는 영향(影響))

  • Kwak, Jung-mun;Oh, Min-Suck;Song, Tae-won
    • Journal of Haehwa Medicine
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    • v.8 no.1
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    • pp.625-641
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    • 1999
  • Aging occurs as a part of maturation as the time progresses which manifests in the human body causing morphological and functional degeneration, eventually leading to death. This experimental study was conducted to investigate a herbal formula to fortify the heart with easy clinical applications. Sungshimsan was chosen to study its effects in heart lipid peroxide and metabolic enzyme system in senescence induced rats. After pre-treatment of Sungshimsan for 2 weeks at the dosage of A (100mg/kg), B (250mg/kg), C (350mg/kg), and D (500mg/kg), a lipid peroxide and metabolic enzyme system changes of the heart were meaured in 32 weeks old rats. The following results were obtained in this study: 1. The contents of lipid peroxide was significantly reduced in the experimental groups treated with greater than 2 weeks at 250mg/kg. 2. The enzymatic activity of cytochrome P-450, cytochrome b5, and NADPH-cytochrome P450 reductase were significantly decreased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 3. The activity of glutathione and glutathione S-transferase were significantly increased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 4. The activity of glutathione reductase and glutathione peroxidase were not influenced compared to the control group. 5. The activity of ${\gamma}$-glutamylcystein synthetase was significantly increased in the 250mg/kg, 350mg/kg, and 500mg/kg experimental groups. 6. The activity of enzymes detoxificatioon superoxide dismutase and catalase were not influenced compared to the control group. Summarizing above results suggest that the Sungshimsan has profound effects in the heart lipid peroxide, free radicals, and delaying the heart aging process. Further clinical researches and application can be anticipated on the topic of senility and gerontology.

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The Mechanism of Nephrotoxicity Formation of Methotrexate in STZ-Induced Hyperglycemic Rats (Streptozotocin 유도 당뇨성 흰쥐에서 methotrexate의 신독성 생성기전에 관한 연구)

  • Kim Seok-Hwan;Kim Yeo-Jeong;Lee Joo-Yeon;Kang Hye-Ok;Lee Hang-Woo;Choi Jong-Won
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.259-265
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    • 2006
  • This study is investigated the effect on mechanism of nephrotoxicity formation of methotrexate(MTX) by hyperglycemic by streptozotocin(STZ). MTX was injected daily at two doses of 3, 6 mg/kg for 1 week in STZ-induced hyperglycemic rats. Activities of BUN, creatinine and LDH were significantly increased by treatment with MTX in STZ-induced diabetic group when compared to MTX treatment group in normal rats' Renal lipid peroxide content and activities of cytosolic enzyme were significantly increased in the treatment of MTX in diabetic group. The concentration of glutathione and glutathione biosynthesis enzymes were decreased by treatment with MTX in STZ-induced diabetic group. These results suggest that nephrotoxicity of MTX in STZ-induced hyperglycemic rat was caused by activation of renal metabolizing enzymes in cytosol and decrease of glutathione concentration.

The Therapeutic Effects of Korean Red Ginseng on Carbon Tetrachloride- and Galactosamine-induced Hepatotoxicity in Rats (홍삼의 사염화탄소 및 갈락토사민 유발 간독성에 대한 치료효과)

  • Lee, Chung-Kyu;Han, Yong-Nam;Kim, Na-Young;Choi, Jong-Won
    • Journal of Ginseng Research
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    • v.27 no.1
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    • pp.11-16
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    • 2003
  • In this study, we investgated the effect of Red Ginseng (KRG) on liver damage induced by carbon tetrachloride (CTC) and galactosamine (GalN) in rats using indicator enzymes such as serum alanine/aspartate aminotransferases, sorbital dehydrogenase, lactate dehydrogenase, and ${\gamma}$-glutamyltransferase. Treatment of KRG restored these enzyme activities to near normal level compared to CTC or GalN treatment alone. Treatment of KRG also enhanced hepatic microsomal enzyme system, malondialdehyde formation, and depletion of reduced glutathione content, which were reduced by CTC or GalN. We also found that the decreased activities of glutathione S-transferase and glutathine reductase but not ${\gamma}$-glutamycysteine synthetase after KRG treatment restored to normal level. These results indicate that KRG has potent therapeutic activity against CTC- and GalN-induced hepatotoxicity in rat.

Anti-Proliferative and Anti-Carcinogenic Enzyme-inducing Activities of Delphinidin in Hepatoma Cells

  • Jang, Chan-Ho;Lee, In-Ae;Lim, Hyun-Ae;Kim, Ju-Ryoung;Ha, Young-Ran;Yu, Hoon;Sung, Mi-Kyung;Kim, Jong-Sang
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.641-645
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    • 2007
  • Delphinidin, an aglycone form of anthocyanins, was demonstrated to have anti-carcinogenic potential. The compound at $50\;{\mu}g/mL$ caused a significant increase of quinone reductase activity, an anti-carcinogenic marker enzyme, in mouse hepatoma cell lines (Hepa1c1c7 and BPRc1). Delphinidin enhanced the expression of other detoxifying or antioxidant enzymes including glutathione s-transferase, gamma-glutamylcysteine synthetase, heme oxygenase 1, and glutathione reductase. It suppressed the proliferation of murine hepatoma cells in a dose-dependent manner, with approximately $IC_{50}$ of $70\;{\mu}g/mL$. These results suggest that delphinidin might be useful for cancer prevention.

Effect of BOSINHWAN(BSH) Water Extract on Renal Lipid Peroxide Content and Metabolic Enzyme System (노화과정(老化過程)의 흰쥐에서 보신환(補腎丸)이 신장(腎臟)의 대사효소계(代謝酵素系)에 미치는 영향(影響))

  • Sun, Min-sung;Oh, Min-suk;Song, Tae-won
    • Journal of Haehwa Medicine
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    • v.8 no.1
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    • pp.659-674
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    • 1999
  • The experimental studies were carried out in order to prove the effect of BSH water extract on Renal lipid peroxide content and metabolic enzyme system experimental studies about peroxide content, transferase, enzyme activity were carried out. The result were obtained as follows : 1. In the change of lipid peroxide of renal tissue, all group was decreased, more of two weeks was decreased. 2. In the Change of BUN of renal tissue, all group was decreased. 3. In the change of LDH of urine, all group was not significant. 4. In the change of ${\gamma}$-glutamyltransferasde, Xanthine oxidase, Aldehyde oxidase of urine, all group was decreased. 5. In the change of protein-bound SH, nonprotein-bound SH, glutathione, glutathione S-transferase, ${\gamma}$-Glutamylcystein synthetase of renal tissue, all group was increased. From above results, BSH was had significant effects on the senile, so it is expected to clinical application on senility and geratology.

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Construction of Amylolytic Industrial Brewing Yeast Strain with High Glutathione Content for Manufacturing Beer with Improved Anti-Staling Capability and Flavor

  • Wang, Jin-Jing;Wang, Zhao-Yue;He, Xiu-Ping;Zhang, Bo-Run
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1539-1545
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    • 2010
  • In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

The effect of Bobitang(BBT) water extract on spleen metabolic enzyme system as to aging process in rats (노화과정(老化過程)의 흰쥐에서 보비탕(補脾湯)이 비장(脾臟)의 대사효소계(代謝酵素系)에 미치는 영향(影響))

  • Lee, Dong-jun;Oh, Min-suk;Song, Tae-won
    • Journal of Haehwa Medicine
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    • v.8 no.1
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    • pp.689-710
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    • 1999
  • Bobitang(BBT) is one of the most important prescription that has been used in oriental medicine(dongyibogam) for recovering spleen condition. The study was done to evaluate effects of BBT water extract on the spleen lipid peroxide content and metabolic enzyme system changes. After pretreatment of BBT I (100mg/kg), BBT II(250mg/kg), BBT III(350mg/kg), BBT IV(500mg/kg) for 1 week, lipid peroxide content and metabolic enzyme system changes of the spleen was measured in 8 months rats. The results were obtained as follows : 1. The content of spleen lipid peroxide was significantly decreased in all experimental groups as compared with control, and best in BBT III IV treated groups. 2. The activity of spleen superoxide generation was significantly decreased in all experimental groups as compared with control, and best in BBT IV III treated groups. 3. The activity of cytochrome P-450 and aminopyrine demethylase wasn't significant change. 4. The activity of aniline hydroxylase was significantly decreased in BBT IV II treated groups, xanthine oxidase was significantly decreased in all experimental groups, aldehyde oxidase was significantly decreased in BBT IV treated group as compared with control. 5. The activity of antioxidant enzymes as superoxide dismutase, catalase, glutathione peroxidase was significantly increased in all experimental groups as compared with control. 6. The activity of glutathion S-transferase was significantly increased in all experimental groups, the concentration of spleen glutathione was significantly increased in BBT IV treated group as compared with control. 7. The activity of ${\gamma}$ -glutamylcystein synthetase was significantly increased in BBT III IV I treated groups as compared with control, the activity of glutathione reductase wasn't significant change. From the above results, BBT is cosidered to have effect of remove peroxide content and free radical that was made during ageing process. It is expected that treatment of BBT can be applied in future clinical study of delaying the ageing process.

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The effect of Panax notoginseng saponins on oxidative stress induced by PCV2 infection in immune cells: in vitro and in vivo studies

  • Wang, Qiu-Hua;Kuang, Na;Hu, Wen-yue;Yin, Dan;Wei, Ying-Yi;Hu, Ting-Jun
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.61.1-61.16
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    • 2020
  • Background: Panax notoginseng saponins (PNS) are bioactive substances extracted from P. notoginseng that are widely used to treat cardiovascular and cerebrovascular diseases and interstitial diseases. PNS have the functions of scavenging free radicals, anti-inflammation, improving blood supply for tissue and so on. Objectives: The aim of this study was to investigate the effects of PNS on the oxidative stress of immune cells induced by porcine circovirus 2 (PCV2) infection in vitro and in vivo. Methods: Using an oxidative stress model of PCV2 infection in a porcine lung cell line (3D4/2 cells) and mice, the levels of nitric oxide (NO), reactive oxygen species (ROS), total glutathione (T-GSH), reduced glutathione (GSH), and oxidized glutathione (GSSG) and the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthetase (iNOS) were determined to evaluate the regulatory effects of PNS on oxidative stress. Results: PNS treatment significantly reduced the levels of NO and ROS, the content of GSSG and the activities of XOD, MPO, and iNOS (p < 0.05), while significantly increasing GSH and the ratio of GSH/GSSG in infected 3D4/2 cells (p < 0.05).Similarly, in the in vivo study, PNS treatment significantly decreased the level of ROS in spleen lymphocytes of infected mice (p < 0.05), increased the levels of GSH and T-GSH (p < 0.05), significantly decreased the GSSG level (p < 0.05), and decreased the activities of XOD, MPO, and iNOS. Conclusions: PNS could regulate the oxidative stress of immune cells induced by PCV2 infection in vitro and in vivo.