Literature summary for 1.1.3.10 extracted from

Spadiut, O.; Pisanelli, I.; Maischberger, T.; Peterbauer, C.; Gorton, L.; Chaiyen, P.; Haltrich, D.
Engineering of pyranose 2-oxidase: improvement for biofuel cell and food applications through semi-rational protein design (2009), J. Biotechnol., 139, 250-257.

Cloned(Commentary)

Cloned (Comment)	Organism
heterologously expressed in Escherichia coli BL21 Star DE3	Trametes ochracea

Protein Variants

Protein Variants	Comment	Organism
V546C/E542K	mutant, analysis of kinetic parameters	Trametes ochracea
V546C/T169G	mutant, analysis of kinetic parameters	Trametes ochracea
V546C/T169G/L537W	mutant, analysis of kinetic parameters	Trametes ochracea

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.043	-	1,4-benzoquinone	V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.072	-	1,4-benzoquinone	V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.092	-	ferricenium hexafluorophosphate	V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.092	-	ferricenium hexafluorophosphate	mutant enzyme V546C/T169G/L537W, at pH 6.5 and 30°C	Trametes ochracea
0.15	-	ferricenium hexafluorophosphate	V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.15	-	ferricenium hexafluorophosphate	mutant enzyme V546C/E542K, at pH 6.5 and 30°C	Trametes ochracea
0.22	-	ferricenium hexafluorophosphate	V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.22	-	ferricenium hexafluorophosphate	mutant enzyme V546C/T169G/L537W, at pH 6.5 and 30°C	Trametes ochracea
0.28	-	1,4-benzoquinone	V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.31	-	ferricenium hexafluorophosphate	V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.31	-	ferricenium hexafluorophosphate	mutant enzyme V546C/T169G, at pH 6.5 and 30°C	Trametes ochracea
0.37	-	1,4-benzoquinone	V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.4	-	D-galactose	V546C/T169G mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.44	-	D-glucose	V546C/T169G mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.49	-	D-glucose	V546C/T169G/L537W mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.5	-	ferricenium hexafluorophosphate	V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.5	-	ferricenium hexafluorophosphate	mutant enzyme V546C/T169G, at pH 6.5 and 30°C	Trametes ochracea
0.86	-	D-galactose	V546C/T169G/L537W mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
1.06	-	ferricenium hexafluorophosphate	V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
1.18	-	1,4-benzoquinone	V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
1.5	2	1,4-benzoquinone	V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
1.5	2	D-glucose	V546C/E542K mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
24.6	-	D-galactose	V546C/E542K mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea

Organism

Organism	UniProt	Comment	Textmining
Trametes ochracea	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
protein purification by immobilized metal affinity chromatography, ultrafiltration, concentration of enzymes	Trametes ochracea

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.27	-	V546C/T169G/L537W mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.34	-	V546C/T169G/L537W mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.38	-	V546C/T169G mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
0.43	-	V546C/T169G mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
5.54	-	V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
6.13	-	V546C/E542K mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
24.1	-	V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
42.1	-	V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
51.9	-	V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
56.4	-	V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
82.2	-	V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
82.2	-	V546C/E542K mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
103	-	V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
189	-	V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
221	-	V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
246	-	V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
261	-	V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea
664	-	V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2' azinobis(3-ethylbenzthiazolinesulfonic acid) as the chromogen, 30°C, pH 6.5	Trametes ochracea

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
D-galactose + 1,4-benzoquinone	-	Trametes ochracea	2-dehydro-D-galactose + 1,4-hydroquinone	-	?
D-galactose + ferricenium hexafluorophosphate	-	Trametes ochracea	2-dehydro-D-galactose + ferrocenium hexafluorophosphate	-	r
D-galactose + O2	-	Trametes ochracea	2-dehydro-D-galactose + H2O2	-	?
D-glucose + 1,4-benzoquinone	-	Trametes ochracea	2-dehydro-D-glucose + 1,4-hydroquinone	-	?
D-glucose + ferricenium hexafluorophosphate	-	Trametes ochracea	2-dehydro-D-glucose + ferrocenium hexafluorophosphate	-	r
D-glucose + O2	-	Trametes ochracea	2-dehydro-D-glucose + H2O2	-	?

Synonyms

Synonyms	Comment	Organism
P2Ox	-	Trametes ochracea
pyranose 2-Oxidase	-	Trametes ochracea