• Title/Summary/Keyword: Enzyme multilayer

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Enzyme Sensors Modified with Avidin/Biotin Systembased Protein Multilayers

  • Anzai, Jun-Ichi;Du, Xiao-Yan;Hoshi, Tomonori;Suzuki, Yasuhiro;Takeshita, Hiroki;Osa, Tetsuo
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.591-596
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    • 1995
  • Enzyme multilayers composed of avidin and biotin-labeled enzymes were prepared on the surface of electrode, through a strong affinity between avidin and biotin (binding constant: ca $10^{15} M^{-1}$). The enzyme multilayers were useful for the improvement of the performance characteristies of enzyme sensors. The output current of the enzyme sensors depended linearly on the number of enzyme layers deposited. Thus, lactate oxidase (LOx) and alcohol oxidase (AlOx) were deposited after being modified with biotin for constructing enzyme sensors sensitive to L-lactate and ethanol respectively. It was also possible to deposit two different kinds of enzymes successively in a single multilayer. The glucose oxidase (GOx) and ascorbate oxidase (AsOx) were built into a multilayer structure on a Platinum electrode. The GOx, AsOx multilayer-modified electrode was useful for the elimination of ascorbic acid interference of the glucose sensor.

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Biosensing interfaces based on the dendrimer-underlying layer on gold

  • Yun, Hyeon-Cheol;Kim, Hak-Seong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.52-55
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    • 2000
  • Structually organized mono- and multilayers were developed on gold for the catalytic and affinity biosensing using hyper-branched dendrimers. For the catalytic biosensing interface, a new approach to construct a multilayered enzyme film on the electrode surface was developed. The film was prepared by layer-by-layer depositions of dendrimers and periodate-oxidized glucose oxidase. The voltammograms obtained from the GOx/dendrimer multilayered electrodes revealed that bioelectrocatalytic response is directly correlated to the number of deposited bilayers. From the analysis of voltammetric and ellipsometric signals, the coverage of active enzyme per layer during the layering steps was estimated, demonstrating the spatially-ordered multilayer formation. As an extension of the study, dendrimers having various degrees of ferrocenyl modification were prepared and used. The resulting electrodes were electrochemically characterized, and the density of ferrocenyl groups, active enzyme coverage, and sensitivity were estimated. For the affinity-sensing surrface, a biosensor system based on avidin-biotin interaction was developed. As the building block of affinity monolayer, G4 dendrimer having partial ferrocenyl-tethered surface groups was prepared and used. And the biotinylated and electroactive dendritic monolayer was used for the affinity-sensing surface interacting with avidin. Electrochemical characterization of the resulting biosensor was conducted using free enzyme in electrolyte in terms of degree of surface coverage with avidin and subsequent surface shielding.

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Fabrication of Ultrathin Multilayer Films Using Layer-by-Layer Self-Assembly Method and Their Application (Layer-by-Layer 자기조립현상을 이용한 다층초박막의 제조와 응용)

  • 차국헌;조진한
    • Polymer Science and Technology
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    • v.15 no.3
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    • pp.260-273
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    • 2004
  • Layer-by-layer 자기조립방법 (self-assembly method) 이라고 불리는 담지자기조립방법 (dip self-assembly method)을 이용한 다층초박막은 다양한 전기적, 자기적 성질을 갖는 물질뿐만 아니라 DNA 또는 효소 (enzyme)같은 바이오 물질들을 기판의 크기나 형태에 관계없이 각각의 층에 나노미터 두께로 삽입시킬 수 있음에 따라서 초박막안에 우리가 원하는 특정 성질을 부여할 수 있다. (중략)

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Application of Polymer Brush to Enzyme-Multilayered Porous Hollow-Fiber Membrane

  • Kawakita Hidetaka;Uezu Kazuya;Tsuneda Satoshi;Saito Kyoichi;Tamada Masao;Sugo Takanobu
    • Proceedings of the Membrane Society of Korea Conference
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    • 2004.11a
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    • pp.142-145
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    • 2004
  • Anion-exchange porous hollow-fiber membranes with a thickness of about 1.2 mm and a pore size of about $0.30{\mu}m$ were used as a supporting matrix to immobilize cycloisomaltooligosaccharide glucanotransferase (CITase). CITase was immobilized to the membrane via anion-exchange adsorption and by subsequent enzymatic cross-linking with transglutaminase, the amount of which ranged from 3 to 110 mg per g of the membrane. The degree of enzyme multilayer binding was equivalent to 0.3 to 9.8. Dextran, as the substrate, was converted into seven- to nine-glucose-membered cycloisomaltooligosaccharides (CI-7, -8, and -9) at a maxi mum yield of $28\%$ in weight at a space velocity of 10 per hour during the permeation of $2.0(w/w)\%$ dextran solution across the CITase-immobilized porous hollow-fiber membrane.

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Enhancement of Electrochemical Performance of Cathode by Optimizing Laccase-Carbon Nanotubes Layers for Enzymatic Fuel Cells (Laccase-탄소나노튜브 적층을 통한 효소 연료전지의 cathode 성능 향상)

  • Wang, Xue;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.60 no.4
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    • pp.550-556
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    • 2022
  • The performance of enzymatic fuel cells that convert chemical energy contained in various organic molecules such as sugar, alcohol, organic acids, and amino acids into electrical energy is greatly affected by the cathode as well as the anode. This study aimed to develop a laccase-based cathode with high performance. An enzyme composite composed of an laccase, redox mediator, and carbon nanotubes was immobilized on the surface of electrode in multiple layers, and the effect of the number of layers and the presence or absence of carbon nanotubes on electrode performance was investigated. As the number of layers of the enzyme-mediator (Lac-(PVI-Os-dCl)) on the electrode surface increased, the amount of reduction current generated at the electrode increased. The enzyme-carbon nanotube-mediator composite electrode (Lac-SWCNTs-(PVI-Os-dCl)) generated a current 1.7 times greater than that of the Lac-(PVI-Os-dCl). It was found that the largest amount of current (10.1±0.1 µA) was generated in the electrode composed of two layers of Lac-(PVI-Os-dCl) and two layers of Lac-SWCNTs-(PVI-Os-dCl) in the evaluation of electrodes with different ratio of Lac-SWCNTs-(PVI-Os-dCl) and Lac-(PVI-Os-dCl). The maximum power density of the cell using the cathode composed of a single layer of Lac-(PVI-Os-dCl) and the cell using the optimized cathode were 0.46±0.05 and 1.23±0.04 µW/cm2, respectively. In this study, it was demonstrated that the performance of cathode and the enzymatic fuel cell using the same can be improved by optimizing the layers of composites composed of laccase, redox mediator, and carbon nanotubes on the electrode surface.