Application | Comment | Organism |
---|---|---|
synthesis | recombinant Aga2-GOx fusion proteins in the Saccharomyces cerevisiae cell wall can be used as immobilized catalysts for the production of gluconic acid | Aspergillus niger |
Cloned (Comment) | Organism |
---|---|
recombinant expression of mutant enzyme B11 in Saccharomyces cerevisiae strain EBY100 cell wall | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of enzyme mutant B11 with a C-terminal fusion with Saccharomyces cerevisiae Aga2 protein, the fusion proteins display on the surface of yeast EBY100 cells and show 2fold increased activity compared to the wild-type enzyme at pH 5.5 Aga2-GOx fusion proteins in the yeast cell wall can also be used as immobilized catalysts for the production of gluconic acid. The yeast surface display is developed for the directed evolution of antibodies in Saccharomyces cerevisiae, and involves the fusion of antibody variable domains to Aga2p, the adhesion subunit of the yeast agglutinin protein. Aga2p binds via disulfide bonds to the membrane protein Aga1p, which is embedded in the membrane via a glycosylphosphatidylinositol (GPI) anchor. The Aga2-antibody fusion gene is cloned in the vector pCTCON, whereas the Aga1p gene is integrated into the yeast genome, but both are under the control of galactose-inducible promoters. The surface display system is used for the directed evolution of horseradish peroxidase and expression of GOx for applications in biofuel cells. The kcat of the wild-type and B11 fusion enzymes are 1.65fold and 1.30fold lower than of the non-fusion enzymes, respectively, and the Km values of the wild-type and B11 fusion enzymes are 1.52fold and 1.74fold higher than of the non-fusion enzymes, respectively | Aspergillus niger |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Aspergillus niger | |
16 | - |
beta-D-glucose | recombinant enzyme mutant B11, pH 5.5, 25°C | Aspergillus niger | |
22 | - |
beta-D-glucose | recombinant wild-type enzyme, pH 5.5, 25°C | Aspergillus niger | |
27.9 | - |
beta-D-glucose | recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger | |
33.4 | - |
beta-D-glucose | recombinant wild-type enzyme in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
90000 | 130000 | recombinant glycosylated Aga2GOx fusion protein, native PAGE | 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 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant mutant enzyme B11 from Saccharomyces cerevisiae strain EBY100 cell walls by anion exchange chromatography and ultrafiltration | Aspergillus niger |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-D-glucose + O2 | - |
Aspergillus niger | D-glucono-1,5-lactone + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 100000-140000, recombinant glycosylated Aga2-GOx fusion protein, SDS-PAGE, 2 * 74500, about, recombinant Aga2-GOx fusion protein, sequence calculation, 2 * 65000, about, native GOx, sequence calculation | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
GOX | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Aspergillus niger |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
33.3 | - |
beta-D-glucose | recombinant wild-type enzyme in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger | |
54.8 | - |
beta-D-glucose | recombinant wild-type enzyme, pH 5.5, 25°C | Aspergillus niger | |
61.3 | - |
beta-D-glucose | recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger | |
80 | - |
beta-D-glucose | recombinant enzyme mutant B11, pH 5.5, 25°C | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
recombinant mutant B11 | Aspergillus niger |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Aspergillus niger |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.997 | - |
beta-D-glucose | recombinant wild-type enzyme in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger | |
2.2 | - |
beta-D-glucose | recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C | Aspergillus niger | |
2.49 | - |
beta-D-glucose | recombinant wild-type enzyme, pH 5.5, 25°C | Aspergillus niger | |
5 | - |
beta-D-glucose | recombinant enzyme mutant B11, pH 5.5, 25°C | Aspergillus niger |