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Literature summary extracted from

  • Kovacevic, G.; Blazic, M.; Draganic, B.; Ostafe, R.; Gavrovic-Jankulovic, M.; Fischer, R.; Prodanovic, R.
    Cloning, heterologous expression, purification and characterization of M12 mutant of Aspergillus niger glucose oxidase in yeast Pichia pastoris KM71H (2014), Mol. Biotechnol., 56, 305-311 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.3.4 N,N-dimethyl-4-nitrosoaniline a redox mediator Aspergillus niger

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.4 gene gox, recombinant expression of wild-type and M12 mutant enzymes in Pichia pastoris strain KM71H or in Saccharomyces cerevisiae strain InvSc1 Aspergillus niger

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.4 N2Y/K13E/T30V/I94V/K152R site-directed mutagenesis of mutant M12, pH optimum and sugar specificity of M12 mutant of GOx is similar to the wild-type enzyme, while thermostability is slightly decreased. Mutant M12 GOx expressed in Pichia pastoris shows three times higher activity compared to wild-type GOx towards redox mediators like N,N-dimethyl-nitroso-aniline used for glucose strips manufacturing. Mutant M12 GOx remains very specific for glucose but has higher activity for galactose compared to wild-type GOx Aspergillus niger

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.4 11.43
-
 beta-D-glucose recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 13.33
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger
1.1.3.4 18.1
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 22
-
 beta-D-glucose recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 23.19
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 28.26
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.3.4 160000
-
 about Aspergillus niger

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.4 beta-D-glucose + O2 Aspergillus niger
-
 D-glucono-1,5-lactone + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.4 Aspergillus niger P13006
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
1.1.3.4 glycoprotein the enzyme is around 15% glycosylated with mostly mannose-type glycosylation Aspergillus niger

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.3.4 recombinant wild-type and mutant M12 from Pichia pastoris strain KM71H by cross-flow ultrafiltration and anion exchange chromatography Aspergillus niger

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.3.4 additional information activity of wild-type and mutant M12 GOx during fermentation, overview Aspergillus niger
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.4 beta-D-glucose + O2
-
 Aspergillus niger D-glucono-1,5-lactone + H2O2
-
 ?
1.1.3.4 beta-D-glucose + O2 the reaction can be divided into reductive and oxidative step. In the reductive half of the reaction, beta-D-glucose is oxidized to D-glucono-1,5-lactone, subsequently hydrolyzed to gluconic acid, with simultaneous reduction of FAD to FADH2. In the oxidative half of the reaction, FADH2 in GOx is re-oxidized by oxygen to yield H2O2 Aspergillus niger D-glucono-1,5-lactone + H2O2
-
 ?
1.1.3.4 additional information usage of the nitroso-aniline assay for determination of GOx activity Aspergillus niger ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.4 homodimer 2 * 85000, recombinant enzyme, SDS-PAGE Aspergillus niger

Synonyms

EC Number Synonyms Comment Organism
1.1.3.4 GOX
-
 Aspergillus niger

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.3.4 25
-
 assay at Aspergillus niger

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.3.4 60
-
 purified enzyme, residual activity after 10 min is 32.9% for the wild-type enzyme and 14.7% for enzyme mutant M12 Aspergillus niger

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.4 54.8
-
 beta-D-glucose recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 130.2
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 150
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 189.4
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger
1.1.3.4 257.1
-
 beta-D-glucose recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 352
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.4 5.5 7.4 assay at Aspergillus niger

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.3.4 3.5 9 activity range of wild-type and mutant enzymes, profile overview Aspergillus niger

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.4 FAD the active site holds the tightly, non-covalently bound FAD cofactor Aspergillus niger

General Information

EC Number General Information Comment Organism
1.1.3.4 malfunction enzyme mutant M12 GOx (N2Y/K13E/T30V/I94V/K152R) shows a 3fold higher activity compared to the wild type at 5 mM glucose and two times higher activity at 200 mM glucose Aspergillus niger

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.3.4 2.49
-
 beta-D-glucose recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 5.61
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 6.7
-
 beta-D-glucose recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger
1.1.3.4 8.29
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C Aspergillus niger
1.1.3.4 22.48
-
 beta-D-glucose recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C Aspergillus niger
1.1.3.4 26.41
-
 beta-D-glucose recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C Aspergillus niger