• The Korean Journal of Mycology
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• v.12 no.1
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• pp.21-26
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• 1984

Aspergillus awamori which produces glucoamylase was cultivated in the starch-Czapek-­Dox's medium in which sucrose was depleted. A rapid method for identification and assay of glucoamylase produced by the A. awamori in the culture was established by the use of the glucose oxidase. The levels of glucose derived from the breakdown of the starch medium were assayed by using glucose oxidase, which was proved to be effective in the screening of glucoamylase-producing fungi in terms of rapid and simple determination. After the cellulose acetate electrophoresis of the precipita ted culture broth, the glucoamylase band in the gel was contacted with 2% starch solution with glucose oxidase, and then color reaction was occurred. Also this method could be effective to identify rapidly the fungal glucoamylase.

• KSBB Journal
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• v.9 no.1
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• pp.55-62
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• 1994

Glucose oxidase(EC 1.1.3.4) was purified to electrophoretic homogeneity from Aspergillus niger by a combination of ammonium sulfate fractionation, ion exchange chromatography, and ultrafiltration. Two active fractions A and B, of glucose oxidase were obtained from the hydrophobic chromatography on phenyl sepharose CL-4B. The enzyme A and B were glycoproteins with the same denatured molecular weight of 78, 000 and had specific activities of 2, 191 and 1, 273-units/mg proteins, respectively. But the two enzymes showed differences in native molecular weight that was measured by HPLC gel filteration, maximum absorbtion wavelength and isoelectric point. The enzyme A oxidized $\beta$-D-glucose only and was resistant to sodium dodecyl sulfate. Activity optimum was found at $30^{\circ}C$ and pH 3.5. Also the enzyme A was inhibited greatly by $Hg^{2+}$(10mM). The results of chemical modification experiments suggested that cysteine and cystine residues might be involved in the active site of the enzyme A.

• Applied Chemistry for Engineering
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• v.1 no.2
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• pp.107-115
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• 1990

New insulin-releasing system on the basis of the redox reaction of glucose was synthesized by immobilizing insulin through a disulfide bond(5, 5'-dithiobis(2-nitrobenzoic acid) to polymer membrane(poly(methyl methacrylate)) and enzyme(glucose oxidase). The disulfide bonds were cleaved upon oxidation of glucose with glucose dehydrogenase and glucose oxidase, releasing insulin from the membrane and enzyme. Sensitivity to glucose concentration was enhanced by coimmobilization of enzyme cofactors(nicotinamide adenin dinucleotide and flavin adenin dinucleotide) acting as electron mediator(for the membrane device), and further enhanced by direct immobilization of insulin on glucose oxidase(for the protein device). Both systems were specific to glucose, and the released insulin was indistinguishable from native insulin. The biological activity of released insulin was 81% of native insulin.

• Mycobiology
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• v.39 no.1
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• pp.64-66
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• 2011

Most fungi possess several hydrogen peroxide-generating enzymes, glucose oxidase and pyranose oxidase. Pyranose oxidase can use glucose as its substrate to generate hydrogen peroxide. White rot fungi, which degrade diverse recalcitrant compounds, contain lignin-degrading enzymes, and lignin peroxidase and manganese peroxidase require hydrogen peroxide for their enzymatic reactions. In this study, we isolated a cDNA fragment of pyranose oxidase from Phlebia tremellosa using PCR and examined its expression under the degradation conditions of diethylphthalate (DEP). Pyranose oxidase expression was enhanced up to 30% by the addition of DEP, and this result supports the possible involvement of pyranose oxidase in the degradation of recalcitrant compounds.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.6
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• pp.792-796
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• 1993

The synthesis alcohol-oxidase[EC 1.1.3.13] was investigated in the yeasts, Candida boidinii CBS 8106 and C. boidinii CBS 2428, during growth on different carbon sources. Alcohol-oxidase was undetectable in all strains submitted to the test in the mineral salts medium containing 1.0% glucose, but its production was rapidly increased when the carbon source was changed glucose to 1.0% methanol after 24hrs of incubation. When cells were grown on the various carbon sources (glucose, xylose, lactose, glycerol, galactose, saccharose, sorbose, lactic acid or acetic acid), the alcohol-oxidase activity was undetected. These carbon sources together with methanol yielded far better synthesis of alcohol-oxidase than in the case of carbon sources alone. Alcohol-oxidase was active towards alcohol of shorter alkyl-chain length than C5 and unsaturated alcohols. Its affinity for these alcohols decreased with the increasing length of the alkyl-chain. The apparent Km values for the methanol of Candida boidinii CBS 8106 and C. boidinii CBS 2428 were 1.96 and 1.21, respestively.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1979.10a
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• pp.245.2-246
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• 1979

Heat treated whole cell of Aspergillus niger containing glucose oxidase-catalase system was entrapped in gelatin matrix crosslinked by glutaral-dehyde. The reaction characteristics of immobilized enzyme was studied in a fludized reactor. Heat treatment enhanced the stability and improved the properties of micellium for the immobilized process. The immobilized enzyme system showed the maximum activity at $35^{\circ}C$ and at pH 5.5. The optimum substrate concentration was 0.04M glucose. The activity of immobilized glucose oxidase was in proportion to the concentration of dissolved oxygen in reaction mixture as other reaction conditions were fixed. It was also demonstrated that the limiting factor for the activity of the immobilized glucose oxidase was the oxygen diffusion resistance which increases proportionally to the glucose concentration.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.3
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• pp.224-228
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• 1990

Cabbage contained high peroxidase activity among tested plant sources. The cabbage peroxi-dase can replace horseradish peroxidase to assay glucose with glucose oxidase. The amount of glucose can be determined quantitatively by glucose oxidase-cabbage peroxidase. The opti-mum pH and temperature for enzymatic glucose determination by glucose oxidase-cabbage peroxidase were 6.0 and 35-45$^{\circ}C$ respectively. The glucose assay was inhibited by addition of various metal salts such as mercuric chloride lead acetate silver nitrate ammonium molyb-date sodium tunstate and cupric sulfate. The relationship between absorbance and amount of glucose was linear up to 8.33 mM glucose in the assay mixture under the assay conditions.

• Korean Journal of Materials Research
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• v.18 no.12
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• pp.669-672
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• 2008

Well-aligned Zinc oxide (ZnO) nanowires were synthesized on silicon substrates by a carbothermal evaporation method using a mixture of ZnO and graphite powder with Au thin film was used as a catalyst. The XRD results showed that as-prepared product is the hexagonal wurzite ZnO nanostructure and SEM images demonstrated that ZnO nanowires had been grown along the [0001] direction with hexagonal cross section. As-grown ZnO nanowires were coated with glucose oxidase (GOx) for glucose sensing. Glucose converted into gluconic acid by reaction with GOx and two electrons are generated. They transfer into ZnO nanowires due to the electric force between electrons and the positively charged ZnO nanostructures in PBS. Photoluminescence (PL) spectroscopy was employed for investigating the movements of electrons, and the peak PL intensity increased with the glucose concentration and became saturated when the glucose concentration is above 10 mM. These results demonstrate that ZnO nanostructures have potential applications in biosensors.

• 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 $60^{\circ}C$.

• Applied Biological Chemistry
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• v.27 no.3
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• pp.180-186
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• 1984

For the study of glucose oxidase(GOD) and catalase(CAT) coimmobilization system, the enzymes were obtained from Penicillium spp., PS-8, and the strain itself was used as an immobilizing matrix. To separate glucose oxidase and catalase after the ammonium sulfate fractionation of the culture broth, DEAF-cellulose column was used and its activity yield was 54 and 34%, respectively. Both enzymes were immobilized on the cell matrix, followed crosslinking with 2.5% glutaraldehyde for 12hr. In the determination of efficiencies of GOD and CAT of dual, mixed and soluble enzyme systems, the dual immobilized one w-as superior to those of the soluble or mixed ones. In the comparison of pH profiles, the dual and mixed types showed broader maximum pH ranges than the soluble type. Varying CAT/GOD ratio of the dual system, the higher the ratio showed the broader activity profile. In the comparison of apparent $K_m$ of GOD only and CAT/GOD=10, they were $7.1{\times}10^{-2}$ and $5.1{\times}10^{-2}M$. Their activation energies showed 3.98kcal/mole/deg for GOD only and 2.98kcal/mole/deg for CAT/GOD=10.