• 제목/요약/키워드: $H_2O_2$ catalase

검색결과 290건 처리시간 0.026초

Isolation and characterization of Vitreoscilla mutant defective in catalase-peroxidase hydroperoxidase I

  • Kim, Hee-Jung;Moon, Ja-Young;Lee, John-Hwa;Park, Kie-In
    • 한국동물위생학회지
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    • 제30권3호
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    • pp.291-304
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    • 2007
  • Mutants of an obligate aerobic bacterium, Vitreoscilla, that have deficiency in heat-labile catalase-peroxidase hydroperoxidase I (HPI) were created by EMS treatment. The catalase-peroxidase HPI-deficient mutant showed substantially lower peroxidase activity in exponential and mid-stationary phase compared with the wild type strain. In late stationary phase, the mutant exhibited no peroxidase activity. Peroxidase deficiency in the mutant was revealed by polyacrylamide gels stained for peroxidase activity. Characteristically, catalase levels in the mutant increased about 14- and 8-fold during growth in the exponential and stationary phases, respectively, compared to those in the wild type, suggesting a compensatory effect for protection from $H_2O_2$ toxicity. The mutant showed differences in physiology from the wild type: retardation in growth rate and decrease in oxygen consumption. Both the wild type and the catalase-peroxidase HPI-deficient mutant of Vitreoscilla had lower growth rates in media containing increasing $H_2O_2$ concentrations. However, the mutant exhibited an additionally decreased growth rate after 6 to 8 h of growth compared to the wild type. The wild type was resistent up to 20 mM $H_2O_2$, whereas the mutant was very sensitive to high concentrations of exogenous $H_2O_2$. Although elevated catalase levels would provide protection of the bacteria from the deleterious effect of $H_2O_2$, it did not appear to be complete. Cell-free extracts of the mutant showed decreased NADH oxidation rates and higher accumulation of $H_2O_2$ during this oxidation. These results may account for the impaired growth and earlier onset of death phase by the catalase-peroxidase HPI-deficient mutant of Vitreoscilla.

토마토의 성장과정에 따른 카탈리아제 (Catalase) 활성에 관한 연구 (Studies on the Catalase Activities of Tomato (Lycopersicum esculentum) as a Function of Age)

  • 박시원;김덕심
    • 대한화학회지
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    • 제37권12호
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    • pp.1068-1075
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    • 1993
  • 토마토(Lycopersicum esculentum)를 파종하여 1년간 성장시키면서 성장기간에 따른 잎과 뿌리의 catalase 활성 변화를 측정하였다. 뿌리에 있어서의 catalase 활성은 잎의 경우에 비해 전반적으로 매우 미약하였다. 반면에 잎의 catalase 활성은 매우 현저한 변화 양상을 나타냈는데 주로 peroxisomal 분획에 존재하였고 특히 발아 후 2주 이내의 발아초기에 76 ${\mu}mol$/ml/min의 최고 활성은 기점으로 하여 급격히 감소했다가 성장기 4 ∼ 5 개월에 즈음하여 상당히 증가된 활성을 보였으며 11 ∼ 12개월째 노화기에 가장 저조한 활성을 보였다. 이와 같이 성장시기에 따라서 변화하는 catalase 활성에 영향을 미치는 인자로서 첫째 발아기에는 glyoxylate cycle과 $\beta$-oxidation이 일어나는 glyoxisomal 반응 그리고 둘째 발아기 이후에는 광합성 반응이 각각 해당시기에 $H_2O_2$를 발생시키는 반응으로 작용할 것으로 간주된다. 이 외에도 NADPH는 불활성화된 catalase(compound II)를 다시 활성화시켜 해로운 기질인 $H_2O_2$로부터 catalase 보호제로서의 역할을 함으로써 토마토의 생장기간 전체에 걸쳐 작용하는 것으로 나타났다.

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Pickle 제조과정, $H_2O_2$, 저장기간에 따른 refrigerated dill pickle과 brine중의 lipoxygenase, peroxidase, catalase 활성의 변화 (Effects of Dill Pickling Process, $H_2O_2$ and Storage Duration on Lipoxygenase, Peroxidase and Catalase Activities in Cucumber and Brine)

  • 장미진;조일영;이시경
    • Applied Biological Chemistry
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    • 제39권3호
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    • pp.222-226
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    • 1996
  • 제조과정, 과산화물 및 저장기간에 따른 냉장 Dill 피클과 brine에 함유된 lipoxygenase, peroxidase, catalase 효소의 활성 변화를 조사하였다. 오이를 Dill 피클용 brine에 1일 저장시의 lipoxygenase는 거의 비활성화 되었으나, 1일 이후에는 lipoxygenase의 활성은 서서히 감소하였다. 반면, 오이 tissue에 존재하는 peroxidase의 활성은 피클 제조후 4일 저장기간 동안 서서히 감소 현상을 나타내었다. 신선한 오이 tissue에서 추출된 catalase는 높은 활성을 가지나, 오이를 피클용 brine에 담갔을 경우에는 소량의 catalase 활성만 보였다. Brine에 과산화물을 첨가시, tissue에서 추출된 lipoxygenase, peroxidase, catalase 활성은 저장 기간과 과산화물 농도에 따라 지속적인 감소를 보였다.

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Effects of Cysteine on the Inactivation of Bovine Liver Catalase

  • R. Yousefi;A. A. Saboury;M. Ghadermarzi;A. A. Moosavi-Movahedi
    • Bulletin of the Korean Chemical Society
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    • 제21권6호
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    • pp.567-570
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    • 2000
  • Bovine liver catalase was exposed to cysteine, as a natural inactivator metabolize, causing autoxidation-generating $H_2O_2$ continuously. The catalase species concentrations and activity measurement were done by spectrophotometry in phosphate buffer 10mM, pH 6.5, and 27 $^{\circ}C$. The activity of catalase decreased continuously due to the conversion of active ferricatalase species, E-Fe (III), to an inactive enzyme species, E-Fe (IV). This conversion is related to the slow production of $H_2O_2generated$ by autoxidation of cysteine. The free SH-group of cysteine has an essential role in production of $H_2O_2$ and hence inactivation of catalase. NADPH can protect catalase against inactivation due to the conversion of inactive form of E-Fe (IV) to ferricatalase species, E-Fe (III).

$H_2O$$_2$-Catalase처리에 의한 소규모 목장우유의 일시적 보존법의 개발 (제1보) 우유에 있어서 과산화수소의 살균효과 및 안정성 (Development of Temporary Preservation Method for Small Scale Dairy Farm Milk by $H_2O$$_2$ Catalase Treatment (Part 1) Bactericidal Effect of Hydrogen Peroxide and Its Stability in Milk)

  • 박인식;박무영
    • 한국미생물·생명공학회지
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    • 제5권3호
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    • pp.113-118
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    • 1977
  • 미생물에 의해 오염된 목장우유에 과산화수소를 0.01%에서 0.05% 범위내에서 첨가하고 3$0^{\circ}C$에서 16시간동안 보관하면서 생균수, 잔여 과산화수소량, 유산의 생성 등을 측정해 보았다. 시험된 조건하에서 0.01, 0.02, 0.03%의 과산화수소 처리는 우유속의 생균수를 각각 8, 12, 16시간동안 시초의 오염정도 이하로 유지시킬 수 있었다. 그뿐 아니라 처리한 과산화수소의 농도를 0.03% 이하로 제한했을 때는 catalase의 첨가 없이도 과산화수소가 우유속에서 12시간 이내에 완전히 자연 분해되었으며 그 분해시간은 첨가된 과산화수소의 농도의 감소에 따라 단축되었다. 이런 결과를 토대로 삼아 과산화수소의 첨가량을 줄이므로써 목장우유의 일시적 보관을 위한 보다 안전한 과산화수소의 처리법을 논의하였다.

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폐상피세포 장벽에 대한 $H_2O_2$의 영향 (Effect of $H_2O_2$ on Alveolar Epithelial Barrier Properties)

  • 서덕준;조세헌;강창운
    • Tuberculosis and Respiratory Diseases
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    • 제40권3호
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    • pp.236-249
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    • 1993
  • 연구배경 : 정상 폐상피세포에서는 항상 생성되고 있는 활성산소(oxygen radical)에 의한 유해작용에 노출되어 있고, 이들 유해 산소들은 폐기종과 같은 폐질환의 원인 기전으로 생각되고 있다. 본 연구에서는 이 방법을 이용하여 만든 폐상피세포 단일막에서 전기생리학적인 관점에서 물질의 이동지표인 short-circuit current(Isc)와 조직저항(R)에 대한 활성산소의 하나인 $H_2O_2$(hydrogen peroxide)가 어떤 영향을 미치는지를 연구함으로서 세포생리학적 기전을 구명하고자 한다. 방법 : Tissue culture-treated polycarbonant membrane filter 에서 배양시킨 쥐 제 2 형 폐상피세포 배양 단일막에서 $H_2O_2$의 능동적 이온 이동 (Isc) 과 수동적 용질이동에 대한 조직저항(R)에 미치는 효과를 관찰하였다. 배양 제 3 일과 제 4 일째 단일막을 수정된 Ussing chamber에 설치하고 막 양측에 HEPES-buffered Ringer 용액으로 incubation 하였다. 외부에서 0~100 mM 농도의 $H_2O_2$를 apical 또는 basolateral side에 작용시켜 Isc와 R의 변화를 관찰하였다. 폐상피세포 장벽이 외부의 $H_2O_2$에 대하여 방어작용을 가지는 세포내 catalase 활성도를 측정하고, catalase 억제제인 aminotriazol(ATAZ) 20 mM의 효과도 함께 관찰하였다. 결과 : 이 단일막은 형태학적으로 보아서 in vivo 에서의 포유류 제 1 형 폐상피세포 장벽의 특성을 나타내고 세포들 사이는 tight junction을 이루며(조직저항 R: 2,000 ohm-$cm^2$ 이상) sodium ion의 능동적 이동 (Isc: 5 ${\mu}A/cm^2$)을 보였다. $H_2O_2$는 dose-dependent 양식으로 Isc와 R 모두 감소시켰다. Apical side에 작용하는 $H_2O_2$에 있어서는 60분에 50% 억제하는 농도인 $ED_{50}$는 Isc와 R은 약 4mM 이었으나 basolateral side의 경우는 약 0.04mM 로서 그 작용 강도는 apical에 비하여 약 100배 정도 더 컸다. ATAZ 존재시 apical side의 $ED_{50}$는 0.4mM로 감소하였으나 basolateral side의 경우 변화가 없었다. $H_2O_2$의 제거율은 apical 또는 basolateral side 어느 쪽에 존재하든 같았으며, 세포내 catalase 활성도는세포배양기간이 길어짐에 따라 증가함을 보였다. 결론 : 이상의 실험결과는 basolateral side에 작용하는 $H_2O_2$는 세포내 막구성성분 중 basolateral 측에 존재하는 곳에(예, $Na^+,\;K^+$-APTase) 직접 장애를 미칠 것으로 생각된다. 한편 apical side에 작용하는 $H_2O_2$는 막성분에 도달하기 전에 세포내에 존재하는 catalase에 의하여 대부분 그 작용을 잃게 된다. 결론적으로 Isc와 R로 측정된 폐상피세포 장벽의 특성은 $H_2O_2$에 의하여 손상을 받고 apical side 보다는 basolateral side 측정이 더 손상을 잘 받게 된다.

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Effects of Oxidative Stress on Apoptosis and Antioxidant Enzyme Levels

  • Kim, Choonmi;Lee, Ji-Young
    • Biomolecules & Therapeutics
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    • 제4권4호
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    • pp.402-407
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    • 1996
  • Effects of oxidative stress on the induction of apoptosis and the activity of antioxidant enzymes were investigated in HL-60 cells using $H_2O$$_2$and cisplatin which generate oxygen species in the cell. Various concentrations of oxidants were treated to cells and at different incubation time, cells were harvested for assays. Cell viability, morphology by propidium iodide staining and DNA fragmentation by agarose gel electrophoresis were observed to determine whether they induce apoptosis. The activity of antioxidant enzymes such as superoxide dismutase and catalase was also measured to evaluate the cellular response to the oxidative damage. The results are as follows: $H_2O$$_2$ induced apoptosis at 10 $\mu$M after 6h incubation, while it took 12h for cisplatin. Both oxidants induced the superoxide dismutase activity at a tolerable low concentration. However, at a concentration which causes apoptotic cell death, the enzyme level was dropped markedly at first and then recovered to the normal level after which it declined again, probably due to cell death. On the other hand, changes in the activity of catalase were not significant at most concentrations except the statistically significant decrease at 24h after 10 $\mu$M-$H_2O$$_2$treatment. In this study, $H_2O$$_2$- and cisplatintreated cells showed similar results in apoptotic response and enzyme activities, suggesting that anticancer activity of cisplatin may be related, at least in part, to the production of oxygen free radicals.

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Purification and characterization of catalase-3 of deinococcus radiophilus

  • Lee, In-Jeong;Lee, young-Nam
    • Journal of Microbiology
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    • 제33권3호
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    • pp.239-243
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    • 1995
  • Deinococcus radiophilus, an UV resistant bacterium seemed to contain three issoenzymes of catalase. Among them, the samllest and most abundant species in cell-free extract, catalase-3 which also exhibited peroxidase activity was purified to electrophoretic homogeneity (145-fold purification) by chromatographic procedures. Its molecular weight was 155 kDa composed of four 38 kDa subunits. The $K_{m}$ value of catalase-3 for H$\_$2/O$\_$2/ was approximately 0.5 mM. This enzyme showed a typical ferric heme spectrum with maximum absorption at 405 nm. Upon binding to cyanide, the 405 nm peak shifted to 420 nm. Catalase-3 was very sensitive to inhibitors of heme proteins, such as cyanide, azide and hydroxylamine. A ratio of A$\_$405/A$\_$28O/ was 0.5 Catalase-3 was active over a wide range of pH, between pH 7 and 10. The enzyme was rather heat-labile and partially sensitive to edthanol-chloroform treatment, but resistant to 3-amino-1, 2, 4-triazole. Catalase-3 of D. radiophilus, which is a bifunction catalatic peroxidatic enzyme seemed to share certain molecular properties with the typical catalase and the catalase-[roxidase along with its own unique features.

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황해쑥(Artemisia argyi)의 H9 (ATCC HTB176) 세포에 대한 세포독성 및 항산화효소 활성 (Cytotoxicity of Artemisia argyi Extract Against H9 (ATCC HTB 176) Cell and Antioxidant Enzyme Activities)

  • 김경하;정대영;민태진;박시원
    • 약학회지
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    • 제43권5호
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    • pp.598-605
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    • 1999
  • The hot water and mathanol extracts of Artemisia argyi showed considerable cytotoxicities against H9(ATCC HTB 176) cancer cell with IC50 values of $48.6{\;}\mu\textrm{g}/ml$ and $51.9{\;}\mu\textrm{g}/ml$, respectively. These cytotoxicities were found to be dependent on the extract concentrations and culture days. CuZnSOD and MnSOD activities were significantly increased in the cytoplasm and mitochondria fractions of cancer cell, and media in the presence of Artemisia argyi. Such enhanced SOD activities were generally in the range of two to threefolds. In contrast to SOD, catalase and glutathione peroxidase activities were not detected at all. These results suggest that Artemisia argyi have generated $O_2^-$ in the mitochondria and cytoplasm of H9 cancer cell with concurrent induction of CuZnSOD and MnSOD in situ, which dismutate $O_2^-{\}to{\;}H_2O_2$. Without coordinated actions of catalase and/or glutathione peroxidase $H_2O_2$ is easily converted to very toxic OH and these reactive oxygen species together might have induced necrosis and/or apoptosis of H9 cell.

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$H_2O_2$에 의한 신장(腎臟) 세포 손상에 대한 단삼(丹參) 추출물의 방지 효과 (Protective effect of Salviae-radix extraction in $H_2O_2$ induced renal cell injury)

  • 김상범;정지천
    • 대한한의학회지
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    • 제19권1호
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    • pp.38-48
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    • 1998
  • This study was undertaken to determine whether Salviae-radix (SVR) extraction prevents the oxidant-induced cell injury and thereby exerts protective effect against oxidant-induced inhibition of tetraethylammonium uptake (TEA) in renal corticaJ sices. SVR (5%) attenuated $H_2O_2-induced$ inhibition of TEA uptake. $H_2O_2$ increased LDH release and lipid peroxidation in a dose-dependent manner. These changes were prevented by SVR extraction. The protective effect of SVR on LDH release was dose-dependent over the concentration range of 0.1-0.5%, and that on lipid peroxidation over the concentration ranges of 0.05-2%. SVR significantly prevented Hg-induced lipid peroxidation. SVR extraction (0.5%) increased cellular GSH content in normal and $H_2O_2-treated$ tissues. When slices were treated with 100 mM $H_2O_2$, catalase activity was decreased, which was prevented by 0.5% SVR extraction. The activity of glutathione peroxidase but not superoxide dismutase was significantly increased by 0.5% SVR extraction in $H_2O_2-treated$ tissuces. These results suggest that SVR has an antioxidant action and thereby exerts benefical effect against oxidant-induced impairment of membrane transport function. This effect of SVR is attributed to an increase in endogenous antioxidants such as GSH, catalase and glutathione peroxidase.

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