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Literature summary for 1.1.3.4 extracted from
- 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 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| N,N-dimethyl-4-nitrosoaniline | a redox mediator | Aspergillus niger |  |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 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
| Protein Variants | Comment | Organism |
|---|---|---|
| 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]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 11.43 | - |
beta-D-glucose | recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 13.33 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
| 18.1 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 22 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 23.19 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 28.26 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 160000 | - |
about | Aspergillus niger |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| beta-D-glucose + O2 | Aspergillus niger | - |
D-glucono-1,5-lactone + H2O2 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Aspergillus niger | P13006 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | the enzyme is around 15% glycosylated with mostly mannose-type glycosylation | Aspergillus niger |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant M12 from Pichia pastoris strain KM71H by cross-flow ultrafiltration and anion exchange chromatography | Aspergillus niger |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | activity of wild-type and mutant M12 GOx during fermentation, overview | Aspergillus niger | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| beta-D-glucose + O2 | - |
Aspergillus niger | D-glucono-1,5-lactone + H2O2 | - |
? | |
| 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 | - |
? | |
| additional information | usage of the nitroso-aniline assay for determination of GOx activity | Aspergillus niger | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 85000, recombinant enzyme, SDS-PAGE | Aspergillus niger |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GOX | - |
Aspergillus niger |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Aspergillus niger |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 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]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 54.8 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 130.2 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 150 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 189.4 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
| 257.1 | - |
beta-D-glucose | recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 352 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 7.4 | assay at | Aspergillus niger |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 3.5 | 9 | activity range of wild-type and mutant enzymes, profile overview | Aspergillus niger |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | the active site holds the tightly, non-covalently bound FAD cofactor | Aspergillus niger | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| 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]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2.49 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 5.61 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 6.7 | - |
beta-D-glucose | recombinant wild-type enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
| 8.29 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Saccharomyces cerevisiae, pH 5.5, 25°C | Aspergillus niger | |
| 22.48 | - |
beta-D-glucose | recombinant mutant M12 expressed in Pichia pastoris, pH 7.4, 25°C | Aspergillus niger | |
| 26.41 | - |
beta-D-glucose | recombinant mutant M12 enzyme expressed in Pichia pastoris, pH 5.5, 25°C | Aspergillus niger | |
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