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Literature summary extracted from

  • Blazic, M.; Kovacevic, G.; Prodanovic, O.; Ostafe, R.; Gavrovic-Jankulovic, M.; Fischer, R.; Prodanovic, R.
    Yeast surface display for the expression, purification and characterization of wild-type and B11 mutant glucose oxidases (2013), Protein Expr. Purif., 89, 175-180 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.3.4 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(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.4 recombinant expression of mutant enzyme B11 in Saccharomyces cerevisiae strain EBY100 cell wall Aspergillus niger

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.4 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.4 additional information
-
 additional information Michaelis-Menten kinetics Aspergillus niger
1.1.3.4 16
-
 beta-D-glucose recombinant enzyme mutant B11, pH 5.5, 25°C Aspergillus niger
1.1.3.4 22
-
 beta-D-glucose recombinant wild-type enzyme, pH 5.5, 25°C Aspergillus niger
1.1.3.4 27.9
-
 beta-D-glucose recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C Aspergillus niger
1.1.3.4 33.4
-
 beta-D-glucose recombinant wild-type enzyme in fusion with Aga2, 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 90000 130000 recombinant glycosylated Aga2–GOx fusion protein, native PAGE 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
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.3.4 recombinant mutant enzyme B11 from Saccharomyces cerevisiae strain EBY100 cell walls by anion exchange chromatography and ultrafiltration 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
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.4 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

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.4 33.3
-
 beta-D-glucose recombinant wild-type enzyme in fusion with Aga2, pH 5.5, 25°C Aspergillus niger
1.1.3.4 54.8
-
 beta-D-glucose recombinant wild-type enzyme, pH 5.5, 25°C Aspergillus niger
1.1.3.4 61.3
-
 beta-D-glucose recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C Aspergillus niger
1.1.3.4 80
-
 beta-D-glucose recombinant enzyme mutant B11, pH 5.5, 25°C Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.4 5
-
 recombinant mutant B11 Aspergillus niger

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.4 FAD
-
 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 0.997
-
 beta-D-glucose recombinant wild-type enzyme in fusion with Aga2, pH 5.5, 25°C Aspergillus niger
1.1.3.4 2.2
-
 beta-D-glucose recombinant enzyme mutant B11 in fusion with Aga2, pH 5.5, 25°C Aspergillus niger
1.1.3.4 2.49
-
 beta-D-glucose recombinant wild-type enzyme, pH 5.5, 25°C Aspergillus niger
1.1.3.4 5
-
 beta-D-glucose recombinant enzyme mutant B11, pH 5.5, 25°C Aspergillus niger