• 제목/요약/키워드: gshII

검색결과 89건 처리시간 0.025초

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

  • 남용석;이세영
    • 생명과학회지
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    • 제7권4호
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    • pp.253-261
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    • 1997
  • E. coli에서 글루타치온 생산 증가를 위해서 E. coli에서 분리한 gehI과 gshII유전자를 함유하고 있는 여러 재조합 플라스미드를 구성하여 도입하였다.pBR325 벡터에 gehI 유전자를 각각 1-3개를 포함한 재조합 플라스미드 및 gehI과 gehII 유전자를 동시에 갖는 재조합 플라스미드를 구성하였다. 계속적으로 반복된 gehI 유전자가 증폭된 E. colidml $\gamma $-gluramylcysteine synthetase의 효소활성은 삽입된 gehI 유전자의 부에 따라 증가하였다. 구성된 재조합 플라스미드를 함유한 E.coli의 글루타치온 생산능을 accetate kinase반응을 ATP재생계로 사용하여 조사한 결과 반복된 gehI 유전자를 함유한 E.coli의 글루타치온 생산능력은 삽입된 gehI 유전자의 수에 비례한여 증가하였으며, gehI 유전자의 추가적인 도입에 의해 글루타치온 생산능력은 2배 증가하였다. E.coli에서 글루타치온의 효소적 생산은 주로 \gamma $-gluramylcysteine synthetase의 효소활성에 의해 영향을 받았다. 가장 높은 글루타치온 생산능은 pGH501 (pUC8-gsh.I.II.III) 플라스미드를 갖는 균주에서 관찰되었다.

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

  • 남용석;박영인;이세영
    • 한국미생물·생명공학회지
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    • 제19권6호
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    • pp.575-582
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    • 1991
  • 글루타치온 생산증대를 도모하기 위하여 글루타치온 합성에 관여하는 효소들인 GSH-I 및 GSH-II 유전자들 pBR322 벡터에 클로닝하였다. gshI 유전자를 클로닝하기 위하여 GSH-I 효소활성이 결여된 GS903 균주를 분리하였다. E. coli K-12 염색체 DNA로부터 분리된 3.6Kb PstI DNA 절편을 pBR322 벡터에 클로닝하였다. gshII 유전자는 2.2Kb PstI-BamHI DNA 절편내에 존재하며 이 절편을 pUC13 벡터에 클로닝하였다. 플라스미드의 copy number와 gsh 유전자들을 포함하고 있는 삽입 DNA의 크기의 차이에 의한 gsh 유전자들의 발현 정도를 조사하기 위하여 pGH1090, pGH101, pGH200, pGH201, pLF4, pLF6 그리고 pGH300같은 여러 플라스미드를 사용함으로써 gshI 유전자의 발현이 의해 2배이상 증가하였다. 그러나 벡터 플라스미드내에 존재하는 gsh 유전자들을 포함하는 삽입 DNA의 크기의 차이에 의해서는 gsh 유전자들의 발현이 영향을 받지 않았다.

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불로환(不老丸)을 투여한 흰쥐 뇌의 항산화효과에 관한 연구 (Effect of Bulohwan-Decoction on Antioxidant Enzyme in Rat Brain)

  • 박진성;고성규;이선동
    • 대한예방한의학회지
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    • 제5권1호
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    • pp.90-102
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    • 2001
  • Objectives: Resently Oxidative stress of brain was proved the cause of Alzheimer and stroke sequel. It has important significance in prevention and treatment of cerebropathia that Bulnohwan used as formula of senescence delay have antioxidative effect. The purposes of this study is to investigate the effect of Bulnohwan on antioxidant defense systems such as thiobarbituric acid reactive substances(TBARS), Superoxide dismutase (SOD), Catalase (CAT), Glutathione peroxidase (GSH-PX), Glutathione S-transperase (GST), Glutathione (GSH) in rat brain. Method: Sprague - Dawley rats were divided into 3 groups; saline solution administered control group, Bulnohwan extract administered Experimental group I and Bulnohwan adminisrtrated, 40% dietary restricted Experimental group II. Animals were sacrificed at 12 weeks after treatment TBARS, SOD, CAT, GSH-PX, GST and GSH were measured in mts brain. Results: weight of brain was no stastical significance.(p>0.05) TBARS contents were significant decrease in Experimental group I, II. (p<0.001) SOD activity was stastical significance in Experimental group II, whereas it was no stastical significance Experimental group II.(p<0.0001) Catalase activites were significant increase in . (p<0.00l) Glutathione Peroxidase activites were significant increase in Experimental group I,II. (p<0.000l) Glutathione S-transferase activites were significant increase in Experimental group I, II. (p<0.000) However there were no statistical significance each other. Glutathione contents were significant increase in Experimental group I, II. (p<0.00l) Conclusions: According to the above results, it is considered that Bulohwan has antioxidants effect in rat brain. When Bulohwan goes with diet restriction, there has more Antioxidants effect in rat brain. but this study was perfored retrospectively. So more prospective studies about mutual relation of drugs are needed

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MOLECULAR BREEDING OF GLUTATHIONE PRODUCING BACTERIAL STRAINS

  • 남용석
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 1991년도 춘계학술발표대회 논문집
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    • pp.237-242
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    • 1991
  • In order to increase the production of glutathione by maximizing the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were cloned. A gshI gene was cloned onto pBR322 plasmid as 3.6Kb PstI DNA fragment from E. coli K-12 chromosomal DNA. Also gshII gene was cloned onto pUC13 plasmid as 2.2Kb PstI-BamHI DNA fragment. In order to improve the glutathione producing activity more efficiently, various recombinant plasmids containing tandem repeated gshI genes or both genes in various copy number onto the same vector were constructed. E. coli cells harboring pGH501 plasmid (pUC8-gshI$\cdot$I$\cdot$II) showed the highest glutathione synthesizing activity. The conditions for glutathione production with an ATP-generating system such as acetate kinase reaction of E. coli cells or glycolytic pathway of yeast cells were examined using the E. coli cells harboring the pGH501 plasmid. When the acetate kinase reaction of E. coli cells was used as an ATP generating system, 20mM of L-csteine was converted into glutathione with a yield of $100\%$.

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황기(黃耆) 약침액(藥鍼液)의 Glutathione S-transferase 와 NAD(P)H: Quinone Reductase 유도 (Induction of Glutathione S-transferase and NAD(P)H:Quinone Reductase by Astragali Radix Aqua-acupuncture Solution)

  • 류준선;임종국
    • Korean Journal of Acupuncture
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    • 제18권1호
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    • pp.21-26
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    • 2001
  • 발암물질을 무독화시키는 QR 생성 유도를 살펴보기 위하여 황기 약침액 및 열수추출액을 생쥐의 간암세포인 Hepa1c1c7에 처리하여 측정한 결과, 황기 약침액의 농도를 증가시킬수록 많은 QR 생성율을 보였으며, GSH 생성이 증가하였고, GST 생성 또한 증가하였다.

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천궁이 Phase II 효소 유도와 Ornithine Decarboxylase 활성에 미치는 영향 (Effect of Cnidii Rhizoma on Phase II Enzyme and Ornithine Decarboxylase Activities)

  • 손윤희;김미경;조현정;남경수
    • 동의생리병리학회지
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    • 제20권6호
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    • pp.1572-1575
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    • 2006
  • Water extract from Cnidii Rhizoma (CRW) was tested for colon cancer chemopreventive activity by measuring the induction of phase II detoxification enzyme activity [quinone reductase (QR) and glutathione S-transferase (GST)] and glutathion (GSH) levels and ornithine decarboxylase (ODC) activity in cultured human colorectal adenocarcinoma HT-29 cells. CRW inhibited cell proliferation in cultured HT-29 cells. CRW induced QR activity in a dose-dependent manner in a concentration range of 0.1${\sim}$5.0 $mg/m{\ell}$. GST activity was also induced with the treatment of CRW in HT-29 cells. In addition GSH levels was increased with CRW. CRW inhibited ODC activity, a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. These results suggest that CRW has colon cancer chemopreventive activity by increasing phase II enzyme activity and GSH levels and inhibiting ODC activity in vitro.

당뇨 모델쥐의 간관 췌장에서 타우린이 지질과산화물 생성과 글루타티온 의존성 효소의 활성에 미치는 영향 (Effect of Taurine Supplement on the Lipid Peroxide Formation and the Activity of Glutathione-Dependent Enzyme in the Liver and Islet of Diabetic Model Mice)

  • 임은영;김해리
    • 한국식품영양과학회지
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    • 제24권2호
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    • pp.195-201
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    • 1995
  • 당뇨 모델에서 타우린의 보강에 의한 지질과산화물의 생성과 GSH 관련 효소들의 활성에 미치는 영향을 알아보고자 alioxan을 이용한 I형과 KK-mouse에 고열량식이를 이용하여 II형 당뇨를 유도하였다. I형과 II형 각각에 정상대조군, 타우린보강군, 당뇨군, 타우린보강 당뇨군을 두어, 모두 8개 군으로 나누었으며, 타우린의 보강은 7일 동안 5%(w/v) 수준으로 자유로이 마시게 하였다. 간과 췌장에서 malondialdehycel(MDA), gluta-thione peroxidase(GPX), glutathions S-transferase(GST)의 활성을 측정하였다. 간조직에서 지질과산화물의 함량은 I형의 경우 당뇨군에서 매우 증가했고 타우린 보강에 의한 유의적으로 감소한 것을 볼 수 있었으며, II형에서는 타우린보강에 의해 유의적인 차이가 없었다. 췌장도 간과 같은 결과를 나타내었다. GPX의 활성은 간에서 I형 당뇨군이 유의적으로 증가했으나, II형 당뇨군에서는 유의적으로 감소했다. 타우린의 보강에 의해 GPX활성에는 유의적인 차이가 없었으며 췌장에서도 간과 비슷한 결과를 보였다. GST의 경우에도 당뇨 유도에 의한 활성 변화는 있었으나 타우린의 보강에 의한 활성 변화는 보이지 않았다. 이상의 결과들로 미루어 당뇨에 있어 타우린의 항산화작용은 당뇨 모델의 종류에 따라 다르며, GSH 관련 효소들의 활성변화 보다는 I형 당뇨 모델의 간과 췌장에서 지질과산화물의 생성을 억제하는 작용을 하리라고 생각 된다.

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Nitric oxide modulates antioxidant defense and the methylglyoxal detoxification system and reduces salinity-induced damage of wheat seedlings

  • Hasanuzzaman, Mirza;Hossain, Mohammad Anwar;Fujita, Masayuki
    • Plant Biotechnology Reports
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    • 제5권4호
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    • pp.353-365
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    • 2011
  • The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

Regulation of Nrf2 Mediated Phase II Enzymes by Luteolin in human Hepatocyte

  • Park, Chung Mu
    • 대한의생명과학회지
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    • 제20권2호
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    • pp.56-61
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    • 2014
  • This study attempted to confirm the antioxidative potential of luteolin against tert-butyl hydroperoxide (t-BHP) induced oxidative damage and to investigate its molecular mechanism related to glutathione (GSH)-dependent enzymes in HepG2 cells. Treatment with luteolin resulted in attenuation of t-BHP induced generation of reactive oxygen species (ROS) and oxidative stress-mediated cell death. In addition, accelerated expression of GSH-dependent antioxidative enzymes, glutathione peroxidase (GPx) and glutathione reductase (GR), and heme oxygenase (HO)-1, as well as strengthened GSH content was induced by treatment with luteolin, which was in accordance with increased nuclear translocation of nuclear factor-erythroid 2 p45-related factor 2 (Nrf2), a transcription factor for phase 2 enzymes, in a dose-dependent manner. These results suggest that the cytoprotective potential of luteolin against oxidative damage can be attributed to fortified GSH-mediated antioxidative pathway and HO-1 expression through regulation of Nrf2 in HepG2 cells.

한우 난소 내 Angiotensin II의 분포와 이의 첨가가 체외성숙 및 수정에 미치는 영향에 관한 연구 (Localization of Angiotensin II in Korean Bovine Follicles and Its Effects on IVM/IVF of Oocytes)

  • Quen, J. H.;M. H. Lee;S. K. Kim
    • Reproductive and Developmental Biology
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    • 제28권1호
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    • pp.59-63
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    • 2004
  • 1. The concentrations of Ang. II were 7.20.91 ${\times}$ $10^3$ , 3.80.34 ${\times}$ $10^3$, 3.50.30 ${\times}$ $10^3$, 2.80.22 ${\times}$ $10^3$ pg/ml in bovine follicular fluids from 1∼3 mm, 3∼5 mm, 5∼7 mm and 8∼10 m follicles, respectively. The concentrations of Ang. II decreased in follicular fluids from large follicles. 2. When oocytes were cultured in media containing various concentrations of Ang. II, a higher proportion of oocytes developed to MII stage in medium with 100 ng/ml (79.5%) Ang II compare to that without Ang. II (58.8%). When oocytes from different sizes of follicles were separately cultured in media containing 100 ng/ml Ang. II, maturation rates were higher in oocytes from small and medium follicles those from controls. 3. GSH content in oocytes cultured for 24 hrs in TCM-199 medium containing 10 and 100 ng/ml of Ang. II was also higher than that of oocytes cultured in medium containing 0 or 10 ng/ml Ang. II. When oocytes were cultured in media containing 0, 10, 100, 1,000 ng/ml of Ang. II, the concentrations of GSH were 5.1M, 5.5M, 7.2M, 8.7M, respectively. 4. When oocytes were cultured in media containing various concentrations of 10, 100, 1,000 ng/ml Ang. II, in vitro maturation and developmental rates were 84.0%, 90.0%, 78.0% and 28.0%, 36.0%, 20.0%, respectively. When oocytes were cultured with an addition of Ang. II in media, in vitro maturation rates higher than that of their controls (76.0%).