• 제목/요약/키워드: redox enzyme

검색결과 107건 처리시간 0.026초

Polypyrrole-Glucose Oxidase 효소전극의 배위자 크기에 따른 전기 화학적 특성 (Electrochemical Properties of Polypyrrole-Glucose Oxidase Enzyme Electrode Depending on Dopant Size)

  • 김현철;구할본;사공건
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 하계학술대회 논문집
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    • pp.745-748
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    • 2001
  • We synthesized polypyrrole (PPy) by electrolysis of the pyrrole monomer solution containing support electrolyte KCl and/or p-toluene sulfonic acid sodium salt (p-TS). The electrochemical behavior was investigated using cyclic voltammetry and AC impedance. In the case of using electrolyte p-TS, the redox potential was about -0.3 V vs. Ag/AgCl reference electrode, while the potential was about 0 V for using electrolyte KCl. It is considered as the backbone forms a queue effectively by doping p-T S. Therefore, it is possible to be arranged regularly. That leads to improvement in the electron hopping. The AC impedance plot gave a hint of betterment of mass transport. PPy doped with p-TS has improved in mass transport, or diffusion. That is because the PPy doped with p-TS has a good orientation, and is more porous than PPy with KCl.

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포도당 산화효소를 고정화한 Polypyrrole 효소전극의 배위자 변화에 다른 전기화학적 특성 (Electrochemical Properties of Polypyrrole Enzyme Electrode Immobilized Glucose Oxidase with Different Ligand)

  • 김현철;구할본
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 추계학술대회 논문집 Vol.14 No.1
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    • pp.529-532
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    • 2001
  • We synthesized polypyrrole (PPy) by electrolysis of the pyrrole monomer solution containing support electrolyte KCl and/or p-toluene sulfonic acid sodium salt (p-TS). The electrochemical behavior was investigated using cyclic voltammetry and AC impedance. In the case of using electrolyte p-TS, the redox potential was about -0.3 V vs. Ag/ AgCl reference electrode, while the potential was about 0 V for using electrolyte KCl. It is considered as the backbone forms a queue effectively by doping p-TS Therefore, it is possible to be arranged regularly. That leads to improvement in the electron hopping. The AC impedance plot gave a hint of betterment of mass transport. PPy doped with p-TS has improved in mass transport, or diffusion. That is because the PPy doped with p-TS has a good orientation, and is more porous than PPy with KCl.

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포도당 산화효소를 고정화한 Polypyrrole 효소전극의 배위자 변화에 따른 전기화학적 특성 (Electrochemical Properties of Polypyrrole Enzyme Electrode Immobilized Glucose Oxidase with Different Ligand)

  • 김현철;구할본
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 추계학술대회 논문집
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    • pp.529-532
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    • 2001
  • We synthesized polypyrrole (PPy) by electrolysis of the pyrrole monomer solution containing support electrolyte KCl and/or p-toluene sulfonic acid sodium salt (p-TS). The electrochemical behavior was investigated using cyclic voltammetry and AC impedance. In the case of using electrolyte p-75, the redox potential was about -0.3 V vs. Ag/AgCl reference electrode, while the potential was about 0 V for using electrolyte KCl. It is considered as the backbone forms a queue effectively by doping p-TS Therefore, it is possible to be arranged regularly. That leads to improvement in the electron hopping. The AC impedance plot gave a tent of betterment of mass transport. PPy doped with p-TS has improved in mass transport, or diffusion. That is because the PPy doped with p-TS has a good orientation, and is more porous than PPy with KCl.

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Expression of a Glutathione Reductase from Brassica rapa subsp. pekinensis Enhanced Cellular Redox Homeostasis by Modulating Antioxidant Proteins in Escherichia coli

  • Kim, Il-Sup;Shin, Sun-Young;Kim, Young-Saeng;Kim, Hyun-Young;Yoon, Ho-Sung
    • Molecules and Cells
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    • 제28권5호
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    • pp.479-487
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    • 2009
  • Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.

Cytoprotection Against Oxidative Damage by Nrf2-regulated Genes

  • Kwak, Mi-Kyoung;Kensler, Thomas W.
    • Toxicological Research
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    • 제23권3호
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    • pp.207-214
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    • 2007
  • Chronic oxidative stress produced by exposure to environmental chemicals or pathophysiological states can lead animals to aging, carcinogenesis and degenerative diseases. Indirect antioxidative mechanisms, in which natural or synthetic agents are used to coordinately induce the expression of cellular antioxidant capacity, have been shown to protect cells and organisms from oxidative damages. Electrophile and free radical detoxifying enzymes, which were originally identified as the products of genes induced by cancer chemopreventive agents, are members of this protective system. The NFE2 family transcription factor Nrf2 was found to govern expression of these detoxifying enzymes, and screening for Nrf2-regulated genes has identified many gene categories involved in maintaining cellular redox potential and protection from oxidative damage as Nrf2 downstream genes. Further, studies using Nrf2-deficient mice revealed that these mutant mice showed more susceptible phenotypes towards exposure to environmental chemicals/carcinogens and in oxidative stress related disease models. With the finding that cancer chemopreventive efficacy of indirect antioxidants (enzyme inducers) is lost in the absence of Nrf2, a central role of Nrf2 in the antioxidative protective system has been firmly established. Promising results from cancer prevention clinical trials using enzyme inducers propose that pharmacological interventions that modulate Nrf2 can be an effective strategy to protect tissues from oxidative damage.

Application of Polyaniline to an Enzyme-Amplified Electrochemical Immunosensor as an Electroactive Report Molecule

  • Kwon, Seong-Jung;Seo, Myung-Eun;Yang, Hae-Sik;Kim, Sang-Youl;Kwak, Ju-Hyoun
    • Bulletin of the Korean Chemical Society
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    • 제31권11호
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    • pp.3103-3108
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    • 2010
  • Conducting polymers (CPs) are widely used as matrixes for the entrapment of enzymes in analytical chemistry and biosensing devices. However, enzyme-catalyzed polymerization of CPs is rarely used for immunosensing due to the difficulties involved in the quantitative analysis of colloidal CPs in solution phase. In this study, an enzyme-amplified electrocatalytic immunosensor employing a CP as a redox marker has been developed. A polyanionic polymer matrix, $\alpha$-amino-$\omega$-thiol terminated poly(acrylic acid), was employed for precipitation of CP. The acrylic acid group acts as a polyanionic template. The thiol terminus of the polymer was used to produce self-assembled monolayers (SAMs) on Au electrodes and the amine terminus was employed for immobilization of biomolecules. In an enzymeamplified sandwich type immunosensor, the polyaniline (PANI) produced enzymatically is attracted by the electrostatic force of the matrix polymer. The precipitated PANI was characterized by electrochemical methods.

고지방 식이로 유도된 비만흰쥐의 체내 지질패턴 및 항산화효소 활성에 도토리 급여의 효과 (Effects of Acorn Supplementation on Lipid Profiles and Antioxidant Enzyme Activities in High Fat Diet-Induced Obese Rats)

  • 강명화;이지현;이정숙;김주현;정혜경
    • Journal of Nutrition and Health
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    • 제37권3호
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    • pp.169-175
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    • 2004
  • This study was performed to investigate the effect of acorn supplementation on the lipid profile and redox antioxidant enzyme activities in obese rat. Obesity in the rats was induced by feeding diet contained 10% lard and 0.5% cholesterol for 4 week. After 4 weeks, rats were divided into the following 5 groups; high fat diet (Control), high fat diet plus 10% Acorn powder (APlO%), high fat diet plus 20% Acorn powder (AP20%), high fat diet plus 0.2% Acorn extract (AE0.2%), high fat diet plus 0.5% Acorn extract (AE0.5%). Total food intake and food efficiency ratio (FER) was not significantly different by acorn powder and extract supplementation. But, body weight was decreased by 20% acorn powder. Acorn powder and extract supplementation for 4 weeks tend to decrease total cholesterol and triglyceride level on the serum and hepatic tissue. There was no significant difference in hepatic glutathione (GSH) content among all the groups. The hepatic GST activity in acorn supplemented groups was lower than that of control. Glutathione peroxidase and catalase activities were higher in acorn supplemented groups than that of control. Hepatic TBARS levels of experimental groups were also significantly lower than that of control group. Our finding suggest that acorn powders and extract might have potential role for improving lipid profiles and antioxidant enzyme activities in obese rats.

Purification and Characterization of the Laccase Involved in Dye Decolorization by the White-Rot Fungus Marasmius scorodonius

  • Jeon, Sung-Jong;Lim, Su-Jin
    • Journal of Microbiology and Biotechnology
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    • 제27권6호
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    • pp.1120-1127
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    • 2017
  • Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.

Clostridium butyricum 유래 Thiolase의 입체구조규명 연구 (Crystal Structure of Thiolase from Clostridium butyricum)

  • 김언정;김경진
    • 생명과학회지
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    • 제26권3호
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    • pp.353-358
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    • 2016
  • Thiolase는 두 분자의 acetyl-CoA를 중합하여 acetoacetyl-CoA를 생산하는 효소이다. 우리는 Clostridium butyricum 유래 thiolase (CbTHL)를 대장균에서 발현하고, 대량으로 정제하여 결정화에 성공하였다. 성장된 결정을 이용하여 엑스선회절 데이터를 획득하였으며, 3차원 입체구조를 2.0 Å으로 규명하였다. 전체적인 구조는 C. acetobutylicum 유래 thiolase (CaTHL)와 같은 type II biosynthetic thiolase와 매우 유사하다는 것을 확인하였다. CbTHL 구조를 CaTHL/CoA 복합체구조와 겹치기를 함으로써 활성화 잔기와 기질결합에 관여하는 잔기를 밝혀낼 수 있었다. CbTHL의 활성화부위는 3개의 잘 보존된 잔기인 Cys88, His349, Cys379로 구성되어 있으며, 이들 잔기는 각각, 공유결합친핵체, 일반염기, 2번째 친핵체의 역할을 하는 것으로 보인다. CbTHL에서 기질결합은, β-mercaptoethyamine과 pantothenic acid 부위는 타 thiolase와 매우 유사한 방법으로 안정화 되지만, ADP 부분은 타 thiolase와는 달리 매우 특이적인 잔기들을 사용한다. CbTHL 구조에서 가장 특징적인 것은 본 단백질이 가역적 이황화결합을 매개로 산화환원스위치를 통하여 그 활성을 조절한다는 것이다.

전기화학적인 방법을 이용한 탄소나노튜브 전극상의 오스뮴 착물의 고정화 및 혈당센서에 관한 응용 (Electrochemical Immobilization of Osmium Complex onto the Carbon Nano-Tube Electrodes and its Application for Glucose sensor)

  • 최영봉;전원용;김혁한
    • 전기화학회지
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    • 제13권1호
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    • pp.50-56
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    • 2010
  • Multi-wall carbon nano-tube(MWCNT)를 이용해 screen printed carbon electrodes(SPCEs)을 제작하여 혈당센서의 선택성과 감도가 증가됨을 확인 할 수 있었다. 효소촉매반응을 위한 탄소전극으로의 전자이동의 매개체로 8족 금속 원소인 오스뮴을 중심금속으로 일차 아민을 포함하는 피리딘(pyridine) 리간드를 배위시켜 $[Os(dme-bpy)_2(4-aPy)Cl]^{+/2+}$를 합성하였다. 합성된 오스뮴 착물은 순환 전압전류법을 포함한 다양한 전기화학분석방법을 이용하여 전기적 성질을 조사하였다. 전기적 흡착방법을 이용하여 일차 아민을 갖는 착화합물을 전극위에 고정화 하였다. 오스뮴이 고정화된 MWCNT-SPCEs는 일반적인 carbon electrode보다 약 100배가량의 오스뮴이 흡착됨을 확인 할 수 있었다. (${\tau}_0=2.0\;{\times}\;10^{-9}\;mole/cm^2$) 마지막으로 당(Glucose)과 당 분해효소(Glucose Oxidase, GOx)에 의한 촉매반응의 전류를 확인하였고, 당 농도에 따라 선형 변화하는 전류의 양도 확인하였다.