• Journal of the Korean Chemical Society
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• v.37 no.11
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• pp.937-942
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• 1993

Enzyme electrodes for amperometric measurement of ammonia was prepared by immobilization of L-glutamate dehydrogenase on an Immobilon-AV Affinity membrane and attachment to a glassy carbon electrode. Reduced nicotinamide adenine dinucleotide (NADH) was used as the electroactive species. The electrochemical oxidation of NADH was monitored at ＋1.0 volt vs. Ag/AgCl. Response was linear from $4.0\;{\times}\;10^{-5}\;to\;4.0\;{\times}\;10^{-4}$ M. The detection limit was 2.0 ${\times}\;10^{-6}$ M. Response time, the optimum pH and life time of enzyme immobilized membrane were 2 min, pH 7.3∼7.6 (Dulbecco's buffer solution) and about 25 days respectively. When the enzyme electrode was applied to the $NH_4^+$ determination with amperometric method, other physiological materials had no interference.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.169-175
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• 2006

Potentiometric biosensor using flow injection analysis system was developed to determine citrate concentration in foods. Biosensor system consisted of sample injector, peristaltic pump, enzyme reactor, carbonate ion selective solid-state electrode, reference electrode, detector, and recorder. Enzyme reactor was prepared with immobilized citrate lyase and oxaloacetate decarboxylase. Carbonate ions produced through enzyme reactions of citrate were potentiometrically detected by ion selective electrode. Optimum conditions for biosensor system were investigated. Interference effect of major sugars and organic acids was less than 5% on citrate biosensor system. Citrate concentrations in fruit juices were determined by biosensor and gas chromatography. No significant difference was observed between two analytical methods. Results indicate citrate biosensor is useful in determining citrate concentration in foods.

• Journal of Biomedical Engineering Research
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• v.3 no.2
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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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• v.15 no.2
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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.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.15 no.7
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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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• v.25 no.10
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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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• v.31 no.10
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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.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.15 no.7
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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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• v.8 no.2
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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.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.15 no.2
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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.