EC Number | Application | Comment | Organism |
---|---|---|---|
1.1.3.4 | additional information | despite the broad range of applications for glucose oxidase, the effectiveness of glucose oxidase is restricted by the narrow substrate range of this enzyme and susceptibility to H2O2 inactivation | Aspergillus niger |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.3.4 | recombinant expression of His-tagged enzyme in Pichia pastoris strain KM71H | Aspergillus niger |
1.1.99.B3 | recombinant expression of His-tagged mutant enzyme in Pichia pastoris strain KM71H | Sarocladium strictum |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.99.B3 | Y300A | the mutant variant of gluco-oligosaccharide oxidase (GOOX) from Sarocladium strictum shows a broader substrate range and higher H2O2 stability compared to the wild-type enzyme. The mutant Y300A exhibits up to 40times higher activity on all tested sugars except glucose, compared to wild-type. Fusion of the Y300A variant to a family 22 carbohydrate binding module from Clostridium thermocellum (CtCBM22A) nearly doubles its catalytic efficiency on glucose, while retaining significant activity on oligosaccharides. In the presence of 200 mM of H2O2, the recombinant CtCBM22A_Y300A retains 80% of activity on glucose and 100% of activity on cellobiose, the preferred substrate for this enzyme, while the wild-type enzyme retains 60% activity on D-glucose under the same conditions. GOOX variants appear to undergo a different mechanism of inactivation, as a loss of histidine instead of methionine is observed after H2O2 incubation. The addition of CtCBM22A also promotes functional binding of the fusion enzyme to xylan, facilitating its simultaneous purification and immobilization using edible oat spelt xylan, which might benefit the usage of this enzyme preparation in food and baking applications. The presence of CtCBM22A also permits enzyme binding to oat spelt xylans, facilitating simultaneous purification and immobilization of the enzyme, and ancillary fibre enrichment in potential food applications | Sarocladium strictum |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.1.3.4 | H2O2 | loss of 40% activity on D-glucose at 200 mM H2O2 | Aspergillus niger | |
1.1.99.B3 | additional information | mutants Y300D and CtCBM22A_Y300A GOOX variants show no H2O2 inactivation on cellobiose | Sarocladium strictum |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.99.B3 | 0.21 | - |
cellotriose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.25 | - |
cellobiose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.25 | - |
cellobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.25 | - |
cellotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.3 | - |
cellopentaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.31 | - |
cellohexaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 3 | - |
xylohexaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 3.15 | - |
xylotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 4.49 | - |
xylopentaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 4.57 | - |
xylotriose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 5.11 | - |
xylobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 5.4 | - |
D-glucose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 5.84 | - |
xylobiose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 8.1 | - |
D-glucose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 37 | - |
D-xylose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 51.8 | - |
D-xylose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum |
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 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.3.4 | Aspergillus niger | Q9HFQ1 | - |
- |
1.1.99.B3 | Sarocladium strictum | Q6PW77 | - |
- |
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 | the enzyme is highly specific for D-glucose | Aspergillus niger | D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.3.4 | additional information | the enzyme oxidizes the anomeric carbon of beta-D-glucose using molecular oxygen as an electron acceptor, producing H2O2 and D-glucono-delta-lactone, which in the presence of water spontaneously hydrolyzes to gluconic acid. Poor activity with xylose, maltose, cellobiose, cellotetraose, and xylo-oligosaccharides | Aspergillus niger | ? | - |
? | |
1.1.99.B3 | cellobiose + O2 | preferred substrate | Sarocladium strictum | ? + H2O2 | - |
? | |
1.1.99.B3 | cellohexaose + O2 | - |
Sarocladium strictum | Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | cellopentaose + O2 | - |
Sarocladium strictum | Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | cellotetraose + O2 | - |
Sarocladium strictum | Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | cellotriose + O2 | - |
Sarocladium strictum | Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | D-glucose + O2 | - |
Sarocladium strictum | D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | D-xylose + O2 | - |
Sarocladium strictum | D-xylono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | maltose + O2 | - |
Sarocladium strictum | 4-O-alpha-D-glucopyranosyl-D-glucono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | additional information | enzyme GOOX oxidizes the anomeric carbon of carbohydrate substrates through an FAD reduction mechanism. The enzyme displays a comparatively broad substrate profile, ranging from D-xylose, and galactose, to malto-, cello- and xylooligosaccharides. GOOX oxidizes oligosaccharides more effectively than monosaccharides | Sarocladium strictum | ? | - |
? | |
1.1.99.B3 | xylobiose + O2 | - |
Sarocladium strictum | xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | xylohexaose + O2 | - |
Sarocladium strictum | D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | xylooligosaccharide + O2 | - |
Sarocladium strictum | ? + H2O2 | - |
? | |
1.1.99.B3 | xylopentaose + O2 | - |
Sarocladium strictum | D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-Xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2 | - |
? | |
1.1.99.B3 | xylotriose + O2 | - |
Sarocladium strictum | xyl-beta-(1->4)-xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.99.B3 | gluco-oligosaccharide oxidase | - |
Sarocladium strictum |
1.1.99.B3 | GOOX | - |
Sarocladium strictum |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.3.4 | 37 | - |
assay at | Aspergillus niger |
1.1.99.B3 | 37 | - |
assay at | Sarocladium strictum |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.99.B3 | 11.17 | - |
cellotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 11.33 | - |
D-xylose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 13.22 | - |
D-glucose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 13.28 | - |
xylobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 13.72 | - |
cellobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 13.87 | - |
xylotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 15.68 | - |
D-glucose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 16.25 | - |
D-xylose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 16.33 | - |
cellobiose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 16.5 | - |
cellohexaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 17.43 | - |
xylohexaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 18.33 | - |
cellopentaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 18.67 | - |
cellotriose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 19.67 | - |
xylotriose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 19.72 | - |
xylobiose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum | |
1.1.99.B3 | 26.17 | - |
xylopentaose | pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A | Sarocladium strictum |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.3.4 | 5 | - |
assay at | Aspergillus niger |
1.1.99.B3 | 8 | - |
assay at | Sarocladium strictum |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.3.4 | FAD | a flavoenzyme with a tightly but non-covalently bound FAD cofactor | Aspergillus niger | |
1.1.99.B3 | FAD | the FAD cofactor is bi-covalently linked through cysteine and histidine residues | Sarocladium strictum |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.3.4 | evolution | glucose oxidase (GO) belongs to the auxiliary activity family AA3_2 | Aspergillus niger |
1.1.3.4 | malfunction | comparison of the substrate profile and H2O2 inactivation of the wild-type glucose oxidase, EC 1.1.3.4, and the Y300A mutant variant of GOOX, the mutant shows a comparatively broad substrate range along with reduced substrate inhibition compared to glucose oxidase, overview | Aspergillus niger |
1.1.3.4 | additional information | narrow binding pocket of glucose oxidase | Aspergillus niger |
1.1.99.B3 | malfunction | comparison of the substrate profile and H2O2 inactivation of the wild-type glucose oxidase, EC 1.1.3.4, and the Y300A mutant variant of GOOX, the mutant shows a comparatively broad substrate range along with reduced substrate inhibition compared to glucose oxidase, overview | Sarocladium strictum |
1.1.99.B3 | additional information | open active site of GOOX compared to glucose oxidase | Sarocladium strictum |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.99.B3 | 0.25 | - |
cellobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.25 | - |
cellotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 0.45 | - |
D-xylose | pH 8.0, 37°C, recombinant enzyme mutant | Sarocladium strictum | |
1.1.99.B3 | 2.883 | - |
D-glucose | pH 8.0, 37°C, recombinant enzyme mutant | Sarocladium strictum | |
1.1.99.B3 | 3.15 | - |
xylotriose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 5.11 | - |
xylobiose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 8.1 | - |
D-glucose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum | |
1.1.99.B3 | 51.8 | - |
D-xylose | pH 8.0, 37°C, recombinant mutant Y300A | Sarocladium strictum |