Activating Compound | Comment | Organism | Structure |
---|---|---|---|
N,N-dimethyl-4-nitrosoaniline | a redox mediator | Aspergillus niger |
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 | 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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
160000 | - |
about | Aspergillus niger |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | P13006 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the enzyme is around 15% glycosylated with mostly mannose-type glycosylation | Aspergillus niger |
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 | Comment | Organism | Textmining |
---|---|---|---|
additional information | activity of wild-type and mutant M12 GOx during fermentation, overview | Aspergillus niger | - |
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 | Comment | Organism |
---|---|---|
homodimer | 2 * 85000, recombinant enzyme, SDS-PAGE | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
GOX | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Aspergillus niger |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | 7.4 | assay at | Aspergillus niger |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3.5 | 9 | activity range of wild-type and mutant enzymes, profile overview | Aspergillus niger |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | the active site holds the tightly, non-covalently bound FAD cofactor | Aspergillus niger |
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 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 |