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Literature summary for 1.1.99.B3 extracted from

  • Vuong, T.V.; Foumani, M.; MacCormick, B.; Kwan, R.; Master, E.R.
    Direct comparison of gluco-oligosaccharide oxidase variants and glucose oxidase substrate range and H2O2 stability (2016), Sci. Rep., 6, 37356 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged mutant enzyme in Pichia pastoris strain KM71H Sarocladium strictum

Protein Variants

Protein Variants Comment Organism
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

Inhibitors

Inhibitors Comment Organism Structure
additional information mutants Y300D and CtCBM22A_Y300A GOOX variants show no H2O2 inactivation on cellobiose Sarocladium strictum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.21
-
cellotriose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
0.25
-
cellobiose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
0.25
-
cellobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
0.25
-
cellotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
0.3
-
cellopentaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
0.31
-
cellohexaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
3
-
xylohexaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
3.15
-
xylotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
4.49
-
xylopentaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
4.57
-
xylotriose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
5.11
-
xylobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
5.4
-
D-glucose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
5.84
-
xylobiose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
8.1
-
D-glucose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
37
-
D-xylose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
51.8
-
D-xylose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum

Organism

Organism UniProt Comment Textmining
Sarocladium strictum Q6PW77
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cellobiose + O2 preferred substrate Sarocladium strictum ? + H2O2
-
?
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
-
?
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
-
?
cellotetraose + O2
-
Sarocladium strictum Glc-beta-(1->4)-Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2
-
?
cellotriose + O2
-
Sarocladium strictum Glc-beta-(1->4)-Glc-beta-(1->4)-D-glucono-1,5-lactone + H2O2
-
?
D-glucose + O2
-
Sarocladium strictum D-glucono-1,5-lactone + H2O2
-
?
D-xylose + O2
-
Sarocladium strictum D-xylono-1,5-lactone + H2O2
-
?
maltose + O2
-
Sarocladium strictum 4-O-alpha-D-glucopyranosyl-D-glucono-1,5-lactone + H2O2
-
?
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 ?
-
?
xylobiose + O2
-
Sarocladium strictum xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2
-
?
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
-
?
xylooligosaccharide + O2
-
Sarocladium strictum ? + H2O2
-
?
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
-
?
xylotriose + O2
-
Sarocladium strictum xyl-beta-(1->4)-xyl-beta-(1->4)-D-xylono-1,5-lactone + H2O2
-
?

Synonyms

Synonyms Comment Organism
gluco-oligosaccharide oxidase
-
Sarocladium strictum
GOOX
-
Sarocladium strictum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Sarocladium strictum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11.17
-
cellotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
11.33
-
D-xylose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
13.22
-
D-glucose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
13.28
-
xylobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
13.72
-
cellobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
13.87
-
xylotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
15.68
-
D-glucose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
16.25
-
D-xylose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
16.33
-
cellobiose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
16.5
-
cellohexaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
17.43
-
xylohexaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
18.33
-
cellopentaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
18.67
-
cellotriose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
19.67
-
xylotriose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
19.72
-
xylobiose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum
26.17
-
xylopentaose pH 8.0, 37°C, recombinant mutant CtCBM22A_Y300A Sarocladium strictum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Sarocladium strictum

Cofactor

Cofactor Comment Organism Structure
FAD the FAD cofactor is bi-covalently linked through cysteine and histidine residues Sarocladium strictum

General Information

General Information Comment Organism
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
additional information open active site of GOOX compared to glucose oxidase Sarocladium strictum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.25
-
cellobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
0.25
-
cellotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
0.45
-
D-xylose pH 8.0, 37°C, recombinant enzyme mutant Sarocladium strictum
2.883
-
D-glucose pH 8.0, 37°C, recombinant enzyme mutant Sarocladium strictum
3.15
-
xylotriose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
5.11
-
xylobiose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
8.1
-
D-glucose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum
51.8
-
D-xylose pH 8.0, 37°C, recombinant mutant Y300A Sarocladium strictum