• KSBB Journal
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• v.11 no.3
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• pp.270-275
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• 1996

Heterologous expression of glucose oxidase gene using recombinant yeast has been carried out. Polymerase chain reaction was conducted to obtain the gene encoding glucose oxidase from Aspergillus niger and sequence comparison indicated the cloned 1.9kb DNA fragment appeared to be the glucose oxidise structural gene containing a signal sequence for extracellular location. Transforming shuttle vector was constructed with YEp352 to express the cloned glucose oxidase gene under the control of either GAL1 or GAL10 promoter. Plate assay of recombinant yeasts has shown that GAL1 promoter was more effective in yielding glucose oxidise than GAL10 promoter. Among the five different concentrations of galactose tried, 1% galactose showed the highest induction of glucose oxidase. Cellular localization experiment of recombinant enzyme using spheroplast revealed that most of enzymes (80%) were secreted into culture media in contrast to A. niger. There is no difference in heat-stability of recombinant enzyme up to $50^{\circ}C$ compared to the glucose oxidase from A. niger However, a dramatic reduction of enzyme activity was observed in both enzymes at $60^{\circ}C$.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2000.07a
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• pp.909-912
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• 2000

We synthesized polypyrrole (PPy) nanotubules by oxidative polymerization of the pyrrole monomer within the pores of a polycarbonate template. The electrochemical behavior was investigated using cyclic voltammetry. The redox potential was about -0.5 V vs. Ag/AgCl reference electrode, while the potential was about 0 V for PPy film. It is considered as the backbone grows according to the pore wall. Therefore, it is possible to be arranged regularly. That leads to improvement in the electron hopping. By electrochemical doping of glucose oxidase (GOx) on PPy nanotubules, an enzyme electrode has been fabricated. The kinetic parameter of biochemical reaction with glucose was evaluated. The formal Michaelis constant and maximum current calculated by computer were about 11.4 mmol $dm^3$ and 170.85 A respectively. Obviously, an affinity for the substrate and current response of the PPy nanotubules enzyme electrode are rather good, comparing with that of PPy film.

• Biomedical Science Letters
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• v.4 no.2
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• pp.103-112
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• 1998

We have developed a simple and accurate strip test that measures the blood glucose level semiquantitatively by visual observation, or qualitatively by using UltraScan spectrocolorimeter. The strip has solid phase reagents, including glucose oxidase, peroxidase, chromogen, affixed to a plastic support. The strip test is capable of measuring blood glucose level in the range of 0∼800 mg/dl and generating the results within 2 to 3 minutes. Human blood specimens obtained from normal individuals and the diabetic patients were evaluated by the new blood glucose strip and by the kit supplied by other commercial products. The test results exhibit the correlation coefficient of 0.964. The new test strip is proven simple and accurate, and it offers an alternative to the commercially available glucose tests.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.417-422
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• 2003

The onions are considered to be a favorable functional source of beverage because they contain much sugar and various nutrients, and they are juicy vegetable. Recently, consumers have a new trend to take functional foods with health benefits. To meet this need, this study was the basic research to establish a manufacturing process of functional onion beverage by glucose oxidase. Glucose oxidase catalyzes reaction of glucose oxidation and makes generation of gluconic acid. Kinetics of the reaction was also investigated, and maximum glucose consumption rate $(V_{max})$ of $26.1{\times}10^{-2}\;g/L{\cdot}min$ and $K_m$ of 5.84 g/L were obtained. Optimum conditions were obtained when the glucose oxidase catalyzed reaction was carried out at temperature of $25^{\circ}C$, agitation rate of 450 rpm and aeration rate of 4 vvm in a 2.5 L jar fermentor. Finally, the enzyme reactor was 10-times scaled up and a similar glucose oxidation performance was achieved in the scaled-up reactor.

• The Korean Journal of Mycology
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• v.51 no.3
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• pp.205-217
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• 2023

This study aimed to produce novel bioactive compounds from non-pathogenic pigmented wild yeasts. Culture supernatants and cell-free extracts of non-pathogenic pigmented yeast strains were prepared, and their physiological functionalities and enzyme activities were measured. Cell-free extracts from Rhodosporidium paludigenum HHGG35-1 and culture supernatants from Rhodosporidium diobovatum NMD18-1 demonstrated very high antioxidant activity (76.6%) and anti-gout xanthin oxidase inhibitory activity (86.2%), respectively. Maximal production of the antioxidants (76.9%) was obtained when Rh. Paludigenum HHGG35-1 was cultured in a yeasts extract-peptone-dextrose (YPD) medium (pH 6.5) at 30℃ for 24 h. The xanthin oxidase inhibitor was also maximally produced (91.6%) when Rh. Diobovatum NMD18-1 was cultured at 30℃ for 96h in a YPD medium (pH 6.5). Rh. Paludigenum HHGG35-1 was oval in shape and formed ascospre. The Rh.diobovatum NMD18-1 specimen displayed dimensions of 1.6 × 1.6 ㎛ and produced ascospores; however, it did not form pseudomycelium. Both of Rh. Paludigenum HHGG35-1 and Rh. Diobovatum NMD18-1 grew well in a 40%-glucose-containing YPD medium and 10%-NaCl-containing YPD medium.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.133-140
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• 1986

Extracts of CO-autotrophically grown cells of Acinetobacter sp. 1 were shown to use thionin, methylene blue, or 2,6-dichlorophenol-indophenol, but not NAD, NADP, FAD, or FMN, as electron acceptors for the oxidation of CO under strictly anaerobic conditions. The CO dehydrogenase (CO-DH) in the thes bacterium was found to be an inducible enzyme. The enzyme activity was determined by an assay based on the CO-dependent reduction of thionin. Maximal reaction rates were found at pH 7.5 and $60^{\circ}C$, and the Arrhenius plot revealed an activation energy of 6.1 kcal/mol(25.5kJ/mol). THe $K_m$ m/ for CO was $154{\mu}M$. Known metalchelating agents tested had no effects on the CO-DH activity. No divalent cations tested affect the enzyme activity significantly escept $Cu^{2+}$ which suppressed the activity completely. The enzyme was inhibited by glucose and succinate. The same extracts catalyzed oxidation of hydrogen gas and formate with thionin as electron acceptor. The CO-DH of Acinetobacter sp. 1 was to have no immunological relationship with that of Pseudomonas carboxydohydrogena.

• Korean Journal of Food Science and Technology
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• v.48 no.2
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• pp.97-103
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• 2016

This study was conducted for the production of water-soluble dietary fiber (SDF) from buckwheat hulls by using Celluclast or Viscozyme. The functionality of this SDF, including antioxidant activity, glucose- and bile acid-retardation effects in vitro, was measured. SDF yields from cellulose and hemicellulose fractions were 60.5 and 123.7 g/kg dry matter, respectively. Analysis of molecular weight distribution of SDF by using gel chromatography showed that SDF degradation increased with increase in reaction time. The antioxidant activity of SDF obtained by enzymatic hydrolysis was higher than that of dietary fiber without enzyme treatment. SDF showed higher retardation effects on glucose and bile acid than the sample without dietary fiber did. The results of this study suggested that SDF produced from buckwheat hull by enzymatic hydrolysis is a good source of functional food material because of its high antioxidant activity and glucose- and bile acid-retardation effects.

• Journal of the Korean Electrochemical Society
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• v.17 no.1
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• pp.30-36
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• 2014

Redox complexes to transport electrons from enzyme to electrodes are very important part in glucose sensor. Pentacyanoferrate-bound aniline ($Fe(CN)_5$-aminopyridine), was prepared as a potential redox mediator in a glucose dehydrogenase (GDH)-glucose sensor. The synthesized pyridyl-$NH_2$ to pentacyanoferrate was characterized by the electrochemical and spectroscopic methods. A amperometric enzyme-linked electrode was developed based on GDH, which catalyses the oxidation of glucose. Glucose was detected using GDH that was co-immobilized with an $Fe(CN)_5$-aminopyridine and gold nano-particles (AuNPs) on ITO electrodes. The $Fe(CN)_5$-aminopyridine and AuNPs immobilized onto ITO electrodes provided about a two times higher electrochemical response compared to that of a bare ITO electrode. As glucose was catalyzed by wired GDH, the electrical signal was monitored at 0.4 V versus Ag/AgCl by cyclic voltammetry. The anode currents was linearly increased in proportion to the glucose concentration over the 0~10 mM range.

• Korean Chemical Engineering Research
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• v.60 no.1
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• pp.62-69
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• 2022

Graphene has a large surface area to volume ratio and good mechanical and electrical property and biocompatibility. This study described the electrochemical deposition and reduction of graphene oxide on the surface of indium tin oxide (ITO) glass slide and electrochemical characterization of graphen-modified ITO. Cyclic voltammetry was used for the deposition and reduction of graphene oxide. The surface of graphen-coated ITO was characterized using scanning electron microscopy and energy dispesive X-ray spectroscopy. The electrodes were evaluated by performing cyclic voltammetry and electrochemical impedance spectroscopy. The number of cycles and scan rate greatly influenced on the coverage and the degree of reduction of graphene oxide, thus affecting the electrochemical properties of electrodes. Modification of ITO with graphene generated higher current with lower charge transfer resistance at the electrode-electrolyte interface. Glucose oxidase was immobilized on the graphene-modified ITO and has been found to successfully generate electrons by oxidizing glucose.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 1999.11a
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• pp.147-150
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• 1999

In the case of immobilizing of glucose oxidase in organic polymer using electrosynthesis, the glucose oxidase 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 glucose oxidase and the backbone of the polymer. That is mainly due to insulating property and net chain of the glucose oxidase. Since being the case, it is useless to increase in amount of glucose oxidase more than reasonable in the synthetic solution. We establish qualitatively that amount of immobilization can be improved by adding a little ethanol in the synthetic solution. As ethanol was added by 0.1 rnol dm" in the synthetic solution, Michaelis-Menten constants of the resulting enzyme electrode decreased from 30.7 mmol $dm^{-3}$ to about 2 mmol $dm^{-3}$. That suggests increase in affinity of the enzyme electrode for glucose and in amount of the immobilized enzyme.zyme.