Application | Comment | Organism |
---|---|---|
diagnostics | the enzyme is used for the manufacture of glucose biosensors and in particular sensor strips used to measure glucose levels in serum | Aspergillus niger |
industry | in the textile industry, enzyme GOX is used for bio-bleaching and in oral care products as antimicrobial agent | Aspergillus niger |
Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain BY4741 (MATa his3 leu2 met15 ura3) from plasmid pSSP-GOX, the GOX coding sequence is fused to the STA1 signal peptide from Saccharomyces cerevisiae var. diastaticus and is under the control of the galactose-inducible GAL10/CYC1 promoter | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
H172K | site-directed mutagenesis, mutant H172K shows increased thermosensitivity compared to the wild-type enzyme | Aspergillus niger |
H172K/H220D | site-directed mutagenesis, mutant H172K/H220D does not show significant differences in thermal stability but about 70% increased initial activity compared to the wild-type enzyme | Aspergillus niger |
H220D | site-directed mutagenesis, mutant H220D shows increased thermosensitivity and reduced activity compared to the wild-type enzyme | Aspergillus niger |
H447K | site-directed mutagenesis, introduction of two symmetrical, intermolecular salt bridges at the dimer interface, between K447 and D70 | Aspergillus niger |
H447K | site-directed mutagenesis, the shows similar initial activity but higher thermal sensitivity compared to the wild-type enzyme | Aspergillus niger |
L500D | site-directed mutagenesis, inactive mutant | Aspergillus niger |
L569E | site-directed mutagenesis, the mutant shows about 50% increased initial activity compared to the wild-type enzyme | Aspergillus niger |
L569E | site-directed mutagenesis, the thermal stability of the mutant is similar to the wild-type enzyme, but the initial activity is increased compared to the wild-type enzyme | Aspergillus niger |
additional information | usage of a strategy that combined random and rational approaches to isolate uncharacterized mutations of Aspergillus niger glucose oxidase with improved properties. GOX library construction in Saccharomyces cerevisiae and random mutagenesis and screening for mutants with improved thermal stability | Aspergillus niger |
Q124R/L569E | site-directed mutagenesis, the mutation has no significant effect on stability but causes a twofold increase of the enzyme's specific activity | Aspergillus niger |
Q345K | site-directed mutagenesis, introduction of the mutation to create a salt bridge with D177 | Aspergillus niger |
Q345K | site-directed mutagenesis, the mutant shows highly reduced thermal stability and about 50% increased initial activity compared to the wild-type enzyme | Aspergillus niger |
Q469K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Aspergillus niger |
Q469K/L500D | site-directed mutagenesis, the mutant shows strongly reduced activity compared to the wild-type enzyme | Aspergillus niger |
Q469K/L500D | site-directed mutagenesis, the thermal stability of the mutant is similar to the wild-type enzyme, but the initial activity is reduced compared to the wild-type enzyme | Aspergillus niger |
Q90R | site-directed mutagenesis, the mutant shows increased sensitivity to thermal denaturation, with R1 and R2 values 60% and 80% lower than wild-type enzyme respectively | Aspergillus niger |
Q90R/Y509E | site-directed mutagenesis, the mutation does not cause a significant change in the thermal stability of the enzyme, but causes increased enzyme activity compared to the wild-type enzyme | Aspergillus niger |
Q90R/Y509E | site-directed mutagenesis, the mutation introduces a new salt bridge near the interphase of the dimeric protein structure, the mutation does not cause a significant change in the thermal stability of the enzyme, but causes increased enzyme activity compared to the wild-type enzyme | Aspergillus niger |
Q90R/Y509E/T554M | the triple mutant is a glucose oxidase with high stability | Aspergillus niger |
T30S/I94V | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Aspergillus niger |
T30S/I94V | site-directed mutagenesis, a thermoresistant mutant | Aspergillus niger |
T554M | random mutagenesis, the mutation generates a sulfur-pi interaction, the mutant shows 60% reduced activity and 40% increased thermal stability compared to the wild-type enzyme | Aspergillus niger |
Y509E | site-directed mutagenesis, the mutation does not cause a significant change in the thermal stability of the enzyme, but causes increased enzyme activity compared to the wild-type enzyme | Aspergillus niger |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
160000 | - |
- |
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 | - |
? | |
beta-D-glucose + O2 | Aspergillus niger CECT 2775 | - |
D-glucono-1,5-lactone + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | A0A068CB13 | sequence with exchanges at V167T and K282E | - |
Aspergillus niger CECT 2775 | A0A068CB13 | sequence with exchanges at V167T and K282E | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | - |
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 | cofactor FAD is transiently reduced along the reaction mechanism | Aspergillus niger | D-glucono-1,5-lactone + H2O2 | - |
? | |
beta-D-glucose + O2 | - |
Aspergillus niger CECT 2775 | D-glucono-1,5-lactone + H2O2 | - |
? | |
beta-D-glucose + O2 | cofactor FAD is transiently reduced along the reaction mechanism | Aspergillus niger CECT 2775 | D-glucono-1,5-lactone + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | each monomer contains two domains, one consists of a five-stranded beta-sheet sandwiched between a three-stranded beta-sheet and three alpha-helices, and the other is composed by a large six-stranded antiparallel beta-sheet supported by six alpha-helices. The two units of the dimer are connected through hydrophobic and hydrophilic contacts, the latter including salt bridges and hydrogen bonds | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
GOX | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Aspergillus niger |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on, the FAD cofactor is not covalently but tightly bound to the enzyme domain, that consists of a five-stranded beta-sheet sandwiched between a three-stranded beta-sheet and three alpha-helices | Aspergillus niger |
General Information | Comment | Organism |
---|---|---|
additional information | identification and analysis of structural motifs of the protein which are critical for its stability | Aspergillus niger |
physiological function | hydrogen peroxide generated by GOX action has anti-microbial effect | Aspergillus niger |