Purification, Properties, and Molecular Features of Glucose Oxidase from Aspergillus niger

Tsuge, Haruhito; Natsuaki, Osamu; Ohashi, Kazuji

doi:10.1093/oxfordjournals.jbchem.a130974

Cited by 140 publications

(79 citation statements)

References 17 publications

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“…Fig.4A represents the effects of pH values on the GOD activity in the presence of glucose, guaiacol and HRP. The GOD activity increased with increasing pH value, and reached a maximum value at pH 5.8, which is close to the optimum pH value (pH5.5) for the native GOD from Aspergillus niger [18][19], indicates that the present GOD assay method does not change the optimum pH value of GOD. The repeatability of the assay was checked by determining the activity of GOD sample repeatly, and the mean value of the GOD activity was calculated to be 114 ± 3U/mg using Eq.3 (see also Fig.4B), with a relative standard deviation (R.S.D) of 2.8% for 31 measurements.…”

Section: Kinetic Parameters Of Godsupporting

confidence: 52%

A Sensitive and Reproducible Spectrophotometric Assay for Glucose Oxidase Activity and Its Application in Conformational Lock

Geng¹,

Li²,

Gao³

et al. 2017

Proceedings of the 2016 International Conference on Biological Engineering and Pharmacy (BEP 2016)

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contributed equally to this work.Abstract-Glucose-oxidase-horseradish peroxidaseguaiacol coupled system was applied for glucose oxidase assay with high sensitivity and prefect reproducibility. The optimum pH value was 5.8. The Michaelis-Menten constant (Km) and Vmax were determined to be 30.0±1.0mM and 2.49±0.08μM/s, respectively, for the assay system at 25º C. The relative standard deviation was 2.8% for 31 times measurements of GOD activity. The GOD assay could be applied in conformational lock study on GOD. Results of bio-thermodynamics suggested that the number of contact area of GOD is two, which is consistent with the computation results by using Ligplot and Pymol.

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Section: Kinetic Parameters Of Godsupporting

confidence: 52%

A Sensitive and Reproducible Spectrophotometric Assay for Glucose Oxidase Activity and Its Application in Conformational Lock

Geng¹,

Li²,

Gao³

et al. 2017

Proceedings of the 2016 International Conference on Biological Engineering and Pharmacy (BEP 2016)

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“…The GOD used in this study is a dimer of two identical subunits with a molecular weight of 160 kDa. 5 The dimer contains two disulfide bonds and two free sulfhydryl groups. Using hydroquinone (HQ)/1,4-benzoquinone (BQ) redox couple as an electron mediator, 6 the inactivation of GOD can be followed by the time-dependent decrease in the HQ/BQ-mediated bioelectrocatalytic current.…”

Section: Introductionmentioning

confidence: 99%

Kinetic Study of the Thermal Inactivation of Glucose Oxidase in the Presence of Denaturant and Stabilizer by Means of Bioelectrocatalysis Method

et al. 2011

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“…The enzyme contains one tightly, noncovalently bound FAD cofactor per monomer and is a homodimer with a molecular mass of 160 kDa, depending on the extent of glycosylation (1). Glucose oxidase from Aspergillus niger is glycosylated by neutral sugars (mostly mannose-like sugars) and by amino sugars (2). Several reports find that the sugar content may vary from 11 up to 30% (3)(4)(5).…”

mentioning

confidence: 99%

Irreversible Thermal Denaturation of Glucose Oxidase from Aspergillus niger Is the Transition to the Denatured State with Residual Structure

Žoldák

Zubrik

Musatov

et al. 2004

Journal of Biological Chemistry

127

105

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Glucose oxidase (GOX; ␤-D-glucose:oxygen oxidoreductase) from Aspergillus niger is a dimeric flavoprotein with a molecular mass of 80 kDa/monomer. Thermal denaturation of glucose oxidase has been studied by absorbance, circular dichroism spectroscopy, viscosimetry, and differential scanning calorimetry. Thermal transition of this homodimeric enzyme is irreversible and, surprisingly, independent of GOX concentration (0.2-5.1 mg/ml). It has an apparent transition temperature of 55.8 ؎ 1.2°C and an activation energy of ϳ280 kJ/mol, calculated from the Lumry-Eyring model. The thermally denatured state of GOX after recooling has the following characteristics. (i) It retains ϳ70% of the native secondary structure ellipticity; (ii) it has a relatively low intrinsic viscosity, 7.5 ml/g; (iii) it binds ANS; (iv) it has a low Stern-Volmer constant of tryptophan quenching; and (v) it forms defined oligomeric (dimers, trimers, tetramers) structures. It is significantly different from chemically denatured (6.67 M GdmHCl) GOX. Both the thermal and the chemical denaturation of GOX cause dissociation of the flavin cofactor; however, only the chemical denaturation is accompanied by dissociation of the homodimeric GOX into monomers. The transition temperature is independent of the protein concentration, and the properties of the thermally denatured protein indicate that thermally denatured GOX is a compact structure, a form of molten globulelike apoenzyme. GOX is thus an exceptional example of a relatively unstable mesophilic dimeric enzyme with residual structure in its thermally denatured state.Glucose oxidase (GOX 1 ; ␤-D-glucose:oxygen oxidoreductase, EC 1.1.3.4) is a flavoenzyme that catalyzes oxidation of ␤-Dglucose by molecular oxygen to ␦-gluconolactone, which subsequently hydrolyzes spontaneously to gluconic acid and hydrogen peroxide. The enzyme contains one tightly, noncovalently bound FAD cofactor per monomer and is a homodimer with a molecular mass of 160 kDa, depending on the extent of glycosylation (1). Glucose oxidase from Aspergillus niger is glycosylated by neutral sugars (mostly mannose-like sugars) and by amino sugars (2). Several reports find that the sugar content may vary from 11 up to 30% (3-5).GOX is of considerable commercial importance. The enzyme has applications in the food and fermentation industry, in the textile industry, and as a molecular diagnostic and analytical tool in medical and environmental monitoring applications (6 -10).The study of GOX and its applications is limited by its conformational instability. A significant effort has been made to increase the stability of GOX. It is known that both the thermal stability and the dynamic properties of the enzyme depend on its redox state (11, 12). Protein glycosylation affects the conformational dynamics of the active site and thus the activity of the enzyme (13). However, the way in which glycosylation affects the stability of GOX is not known. On the other hand, modification of the glucose oxidase surface by artificial long polyethylene-glycol ...

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Purification, Properties, and Molecular Features of Glucose Oxidase from Aspergillus niger

Abstract: 4. FHD (flavin-hypoxanthine dinucleotide) has coenzymatic activity equal to that of FAD.

Cited by 140 publications

References 17 publications

A Sensitive and Reproducible Spectrophotometric Assay for Glucose Oxidase Activity and Its Application in Conformational Lock

A Sensitive and Reproducible Spectrophotometric Assay for Glucose Oxidase Activity and Its Application in Conformational Lock

Kinetic Study of the Thermal Inactivation of Glucose Oxidase in the Presence of Denaturant and Stabilizer by Means of Bioelectrocatalysis Method

Irreversible Thermal Denaturation of Glucose Oxidase from Aspergillus niger Is the Transition to the Denatured State with Residual Structure

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