Crystal Structure of Glucose Oxidase from Aspergillus niger Refined at 2·3 Å Reslution

Hecht, Hans‐Jürgen; Kalisz, Henryk M.; Hendle, J.; Schmid, Rolf D.; Schomburg, Dietmar

doi:10.1006/jmbi.1993.1015

Cited by 646 publications

(527 citation statements)

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“…Members of this subfamily include choline dehydrogenase {Eseherichia coli), glucose dehydrogenase (Drosophila melanogasterL alcohol dehydrogenase (Pseudomonas oleovoraHs), nlethanol oxidase (Han.vemtla poO'morpha and Pichia pastoris), glucose oxidase (Aspergillu,s ni~er) and cholesterol oxidase from Brevihacterium slerolicum. To date, three-dimensional structures have been reported l\~r the two latter enzymes [30,31]. The GMC oxidoreductases all carry FAD as a col'actor, but in contrast to CDH, they do not contain a heine group.…”

Section: Resultsmentioning

confidence: 99%

Cloning and characterization of a cDNA encoding a cellobiose dehydrogenase from the white rot fungus Phanerochaete chrysosporium

et al. 1995

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The eDNA of cellobiose dehydrogenase (CDH) fromPhanerochaete chrysosporium has been cloned and sequenced.The 5' end was obtained by PCR amplification. The cDNA contains 2310 translated bases excluding the poly(A) tail. The deduced mature protein contains 770 amino acid residues and is preceded by a 18 residue long signal peptide. The regions of the amino acid sequence corresponding to the heme and FAD domains of CDH were identified as well as the nucleotide-binding motif, the disulfide pairing and a methionine residue chelating the heme iron. No homologous sequences were found for the heme domain, however, the FAD domain appears to be distantly related to the GMC nxidoreductase family.

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Section: Resultsmentioning

confidence: 99%

Cloning and characterization of a cDNA encoding a cellobiose dehydrogenase from the white rot fungus Phanerochaete chrysosporium

et al. 1995

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“…The 3D structures of GOX and DAOX show topological similarities with regard to the FAD-binding pocket [3,5,19]. Also AO, which displays sequence homology with GOX (approximately 25% identity), may have a similar structure [14].…”

Section: Discussionmentioning

confidence: 98%

“…Also AO, which displays sequence homology with GOX (approximately 25% identity), may have a similar structure [14]. According to the 3D structure of GOX the FAD molecule is embedded in a narrow channel of the protein matrix and forms a number of hydrogen bonds and salt bridges with a number of protein side chains [19]. The network of non-covalent chemical bonds increases the polarization of the functional groups involved in the FAD-binding and makes them more susceptible to the attack of nucleophilic ions like cyanide, cyanate and thiocyanate.…”

Section: Discussionmentioning

confidence: 99%

“…These reagents are even capable of chemical reactions with amino, sulfhydryl, imidazole or phenolic hydroxyl groups or with phosphate anions of the FAD co-factor [20]. In GOX several residues containing such side chains were found to interact with FAD and are therefore likely candidates to react with the nucleophilic reagents [19]. Apparently, the non-covalent bonds involved in the FAD-binding are destabilized during incubation of the proteins with KCN, NaSCN or KCNO resulting in inactivation of the enzymes, but not in dissociation of the co-factor from the proteins.…”

Section: Discussionmentioning

confidence: 99%

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Deflavination of flavo-oxidases by nucleophilic reagents

Zlateva

Boteva

Filippi

et al. 2001

Biochimica et Biophysica Acta (BBA) - Protein Structure and Mol

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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).…”

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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

125

103

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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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Crystal Structure of Glucose Oxidase from Aspergillus niger Refined at 2·3 Å Reslution

Cited by 646 publications

References 0 publications

Cloning and characterization of a cDNA encoding a cellobiose dehydrogenase from the white rot fungus Phanerochaete chrysosporium

Cloning and characterization of a cDNA encoding a cellobiose dehydrogenase from the white rot fungus Phanerochaete chrysosporium

Deflavination of flavo-oxidases by nucleophilic reagents

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

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