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Literature summary extracted from

  • Marin-Navarro, J.; Roupain, N.; Talens-Perales, D.; Polaina, J.
    Identification and structural analysis of amino acid substitutions that increase the stability and activity of Aspergillus niger glucose oxidase (2015), PLoS ONE, 10, e0144289 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.3.4 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
1.1.3.4 industry in the textile industry, enzyme GOX is used for bio-bleaching and in oral care products as antimicrobial agent Aspergillus niger

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.4 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

EC Number Protein Variants Comment Organism
1.1.3.4 H172K site-directed mutagenesis, mutant H172K shows increased thermosensitivity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 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
1.1.3.4 H220D site-directed mutagenesis, mutant H220D shows increased thermosensitivity and reduced activity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 H447K site-directed mutagenesis, introduction of two symmetrical, intermolecular salt bridges at the dimer interface, between K447 and D70 Aspergillus niger
1.1.3.4 H447K site-directed mutagenesis, the shows similar initial activity but higher thermal sensitivity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 L500D site-directed mutagenesis, inactive mutant Aspergillus niger
1.1.3.4 L569E site-directed mutagenesis, the mutant shows about 50% increased initial activity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 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
1.1.3.4 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
1.1.3.4 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
1.1.3.4 Q345K site-directed mutagenesis, introduction of the mutation to create a salt bridge with D177 Aspergillus niger
1.1.3.4 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
1.1.3.4 Q469K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 Q469K/L500D site-directed mutagenesis, the mutant shows strongly reduced activity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 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
1.1.3.4 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
1.1.3.4 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
1.1.3.4 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
1.1.3.4 Q90R/Y509E/T554M the triple mutant is a glucose oxidase with high stability Aspergillus niger
1.1.3.4 T30S/I94V site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Aspergillus niger
1.1.3.4 T30S/I94V site-directed mutagenesis, a thermoresistant mutant Aspergillus niger
1.1.3.4 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
1.1.3.4 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.3.4 160000
-
-
 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
-
 ?
1.1.3.4 beta-D-glucose + O2 Aspergillus niger CECT 2775
-
 D-glucono-1,5-lactone + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.4 Aspergillus niger A0A068CB13 sequence with exchanges at V167T and K282E
-
1.1.3.4 Aspergillus niger CECT 2775 A0A068CB13 sequence with exchanges at V167T and K282E
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
1.1.3.4 glycoprotein
-
 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
-
 ?
1.1.3.4 beta-D-glucose + O2 cofactor FAD is transiently reduced along the reaction mechanism Aspergillus niger D-glucono-1,5-lactone + H2O2
-
 ?
1.1.3.4 beta-D-glucose + O2
-
 Aspergillus niger CECT 2775 D-glucono-1,5-lactone + H2O2
-
 ?
1.1.3.4 beta-D-glucose + O2 cofactor FAD is transiently reduced along the reaction mechanism Aspergillus niger CECT 2775 D-glucono-1,5-lactone + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.4 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

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 37
-
 assay at Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.4 6
-
 assay at Aspergillus niger

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.4 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

EC Number General Information Comment Organism
1.1.3.4 additional information identification and analysis of structural motifs of the protein which are critical for its stability Aspergillus niger
1.1.3.4 physiological function hydrogen peroxide generated by GOX action has anti-microbial effect Aspergillus niger