• 대한화학회지
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• 제37권11호
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• pp.937-942
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• 1993

Immobilon-AV Affinity membrane에 L-glutamate dehydrogenase를 고정하여 유리질 탄소전극에 부착시킨 전극을 사용하여 암모니아를 전류법으로 정량하였다. 이때 환원형의 NADH가 $NAD^+$로 산화될 때 전류를 +1.0 volt vs. Ag/AgCl에서 측정하였다. 효소 고정화된 membrane을 부착시킨 전극의 감응 특성은 다음과 같다. 곧 직선 감응 농도 범위는 $4.0\;{\times}\;10^{-5}\;{\sim}\;4.0\;{\times}\;10^{-4}$ M이었으며, 정량한계는 2.0 ${\times}\;10^{-6}$ M이었다. 또한 감응 시간은 2분이었으며 효소 고정화된 막의 최적 pH와 수명은 각각 pH 7.3∼7.6 (Dulbecco's buffer 용액)과 25일이었다. 그리고 다른 생리활성 물질의 방해는 없었다.

• 한국식품과학회지
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• 제38권2호
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• pp.169-175
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• 2006

Citrate Iyase(CL)와 oxaloacetate decarboxylase(OD)를 고정화하여 효소 반응기를 제작하고 효소반응으로 생성된 carbonate ion을 ion selective electrode를 이용한 전위차법 FIA system으로 식품에함유된 구연산의 농도를 측정하였다. 최적 센서 시스템의 조건은 CL과 OD을 같이 고정화시키는 방법으로 CL과 OD의 양이 각각 10 units, 60 units, 담체량은 0.3 g, carrier buffer 1(0.1 MTris buffer)은 pH 7.5, carrier buffer의 유속은 12 mL/hr으로 결정되었다. 최적 상태에서의 표준검량곡선은 $10^{-1}\;M-10^{-3}\;M$ 범위에서 직선적인 상관관계$(r^2=0.9986)$를 나타내었다. 당류와 유기산류에 대한 센서 시스템에 대한 방해효과를 측정한 결과 당류는 거의 저해효과를 보이지 않았으며 유기산의 경우도 저해 효과가 5% 미만으로 이들 물질에 의해 감응도가 크게 방해받지 않은 것으로 나타났다. 구연산 바이오센서와 GC를 사용하여 식품 시료에 함유된 구연산의 농도를 분석한 결과 두 방법간에 유의적인 차이가 없는 것으로 나타나 본 연구에서 구축한 구연산 바이오센서 시스템은 식품에 함유된 구연산 분석시 신뢰성 있는 결과를 주는 것으로 보여졌다.

• 대한의용생체공학회:의공학회지
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• 제3권2호
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• pp.95-100
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• 1982

Glucose oxidase from A. niger was entrapped in polyacrylamide gel which was used in the enzyme electrode for glucose analysis. The electrode was assembled by placing the gel between the membranes on the surface of a Clark type electrode. In order to make it possible to analyze the experimental results later, the stagnation flow was adopted wheree the governing fluid mechanics were well known. The current increased with the increase concentration in the bulk below a certain level of glucose concentration beyond which no more current increase was observed. This is probably due to the diffusion limitation of oxygen from the bulk solution. Also the current increased witll the enzyme loading in the gel, but the linearity between the current and the glucose concentration was rather limited to a narrow range. Flow rate was found to be very important, which means that film diffusion is very important under the flow rate of 5cm/sec. As a conclusion, enzyme loading, gel layer thickness, stirring speed and bulk concentration of glucose were found to be most improtant parameters in yielding a linar current reponse with respect to the bulk glucose concentration.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.234-238
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• 2005

Glucose oxidase was immobilized on the carboxylated multi-wall carbon nanotubes (MWNT-COOHs) in the presence of a coulping reagent, 1-ethy1-3-(3-dimethylaminopropy1) carbodiimide. Significant amounts of glucose oxidase were also immobilized on MWNT-COOHs without the coupling reagent. Various conditions for the immobilization of glucose oxidase were optimized. Optimal pH for the maximal activity of the immobilized glucose oxidase shifted to 7 from the optimal pH of 6 for the maximal activity of free enzyme due to the carboxy1 groups on the surface of MWNT-COOHs. An electrode of graphite rod with a diameter of 6 mm was fabricated using the immobilized glucose oxidase. The cyclic voltammetry study of the enzyme electrode revealed that the oxidation of glucose and subsequent transfer of electrons from the oxidation of glucose to the electrode were possible by the immobilized glucose oxidase without a mediator, implying that the enzyme electrode can be utilized for the development of biofuel cells.

• 한국전기전자재료학회논문지
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• 제15권7호
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• pp.621-626
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• 2002

In the case of immobilizing of glucose oxidase (GOx) in polypyrrole (PPy) conducting polymer using electrosynthesis, the GOx obstructs charge transfer and mass transport during the film growth. This may lead to short chained polymer and/or make charge-coupling weak between the GOx and the backbone of the PPy. That is mainly due to insulating property and net chain of the GOx. Since being the case, it is useless to increase in amount of GOx mere than reasonable in the synthetic solution. We improved the amount of immobilized GOx into the PPy by adding a little ethanol in the synthetic solution without any more amount of GOx in the solution. We electrochemically analyzed an improvement in the immobilizing event. For the glucose sensing, when ethanol was added by 0.1 mol $dm^{-3}$ in the synthetic solution, the Michaelis constant of the resulting enzyme electrode was about 32 mmol $dm^{-3}$ and maximum current was about $146\mu A$.

• Bulletin of the Korean Chemical Society
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• 제25권10호
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• pp.1499-1502
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• 2004

An amperometric biosensor based on carbon paste electrodes (CPEs) for the determination of urea was constructed by enzyme (urease/GL-DH)-modified method. Urea was hydrolyzed to ${NH_4}^+$ by catalyzing urease onto the enzyme-modified electrode surface in sample solution. In the presence of ${\alpha}$-ketoglutarate and reduced nicotinamide adenine dinucleotide(NADH), a liberated ${NH_4}^+$ produce to L-glutamate and $NAD^+$ by Lglutamate dehydrogenase (GL-DH). After the chemical reaction was proceeded, the electrochemical reaction was occurred that an excess of the NADH was oxidized to $NAD^+$. The oxidation current of NADH was monitored at +1.10 volt vs. Ag/AgCl. An optimum conditions of biosensor were investigated: The optimum pH range for catalyzed hydrolysis reaction of urea was pH 7.0-7.4. The linear response range and detection limit were $2.0\;{\times}\;10^{-5}{\sim}2.0\;{\times}\;10^{-4}M\;and\;5.0\;{\times}\;10^{-6}M$, respectively. Another physiological species did not interfere, except L-ascorbic acid.

• Bulletin of the Korean Chemical Society
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• 제31권10호
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• pp.2913-2917
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• 2010

When rubber dissolved in toluene was used as a binding material of graphite powder, the mechanical robustness of the carbon paste was guaranteed by the fast volatility of the solvent immediately after electrode construction. This characteristic of the rubber solution met qualifications for practical use of carbon paste electrodes and enabled the design of a new enzyme electrode bound with EPDM. In order to confirm whether the electrode shows quantitative electrochemical behaviors or not, its kinetic parameters, e. g. the symmetry factor (0.2), the exchange current density ($3.66\;{\mu}A/cm^2$), the capacity of the double layer ($2.0{\times}10^{-5}\;F$), the Michaelis constant ($4.39{\times}10^{-3}\;M$), the diffusion coefficient of substrate ($2.58{\times}10^{-12}\;cm^2/sec$), the time constant (0.018 sec) and other factors were investigated.

• 한국전기전자재료학회논문지
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• 제15권7호
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• pp.615-620
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• 2002

In the case of immobilizing of glucose oxidase into polypyrrole (PPy) using electrosynthesis, the glucose oxidise (GOx) forms a coordinate bond with the polymers backbone. However, because of intrinsic insulation and net-chain of the enzyme, the charge transfer and mass transport are obstructed during the film growth. Therefore, the film growth is dull. We synthesized enzyme electrodes by electropolymerization added some organic solvent, such as ethanol and tetrahydrofuran (THF). The formative seeds of film growth was delayed by adding ethanol. The delay was induced by radical transfer between ethanol and pyrrole monomer. The radical transfer reactions shared the contribution of dopants between electrolyte anion and GOx polyanion. This led to increase amount of immobilized the enzyme in PPy. For the UV absorption spectra of synthetic solution before synthesis and after, in the case of ethanol added, the optical density was slightly decreased for the GOx peaks. It suggests amount of GOx in the solution was decreased and amount of GOx in the film was increased.

• Journal of Electrochemical Science and Technology
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• 제8권2호
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• pp.87-95
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• 2017

Enzymatic fuel cells are promising low cost, compact and flexible energy resources. The basis of enzymatic fuel cells is transfer of electron from enzyme to the electrode surface and vice versa. Electron transfer is done either by direct or mediated electron transfer (DET/MET), each one having its own advantages and disadvantages. In this study, the DET and MET of laccase-based biocathodes are compared with each other. The DET of laccase enzyme has been studied using two methods; assemble of needle-like carbon nanotubes (CNTs) on the electrode, and CNTs/Nafion polymer. MET of laccase enzyme also is done by use of ceramic electrode containing, ABTS (2,2'-azino-bis [3-ethylbenzthiazoline-6-sulphonic acid]) /sol-gel. Cyclic voltammetric results of DET showed a pair of well-defined redox peaks at $200{\mu}A$ and $170{\mu}A$ in a solution containing 5and $10{\mu}M$ o-dianisidine as a substrate for needle-like assembled CNTs and CNTs-Nafion composite respectively. In MET method using sol-gel/ABTS, the maximum redox peak was $14{\mu}A$ in the presence of 15 M solution o-dianisidine as substrate. The cyclic voltammetric results showed that laccase immobilization on needle-like assembled CNTs or CNTs-Nafion is more efficient than the sol-gel/ABTS electrode. Therefore, the expressed methods can be used to fabricate biocathode of biofuel cells or laccase based biosensors.

• 한국전기전자재료학회논문지
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• 제15권2호
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• pp.141-146
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• 2002

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 oxidation potential of the PPy was about -02 V vs Ag/AgCl reference electrode, while the potential was about 0 V for using electrolyte KCl. The falloff of the oxidation potential gave a sign of an improvement in the electron hopoing mechanism on the backbone. The AC impedance plot gave a hint of betterment of mass transport. PPy doped with p-TS improved in mass transport or diffusion. That was because the PPy doped with p-TS was more porous than PPy with KCl. We attained an effect of good kinetic parameters, in the case of PP-GOx enzyme electrodes doped with p-TS, which were determined by 58 mmol dm$\^$-3/ for apparent Michaelis constant and by 581 ㎂ for maximum current respectively.