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Information on EC 1.1.3.10 - pyranose oxidase and Organism(s) Trametes ochracea and UniProt Accession Q7ZA32
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1.1.3.10 pyranose oxidaseIUBMB Comments
A flavoprotein (FAD). Also oxidizes D-xylose, L-sorbose and D-glucono-1,5-lactone, which have the same ring conformation and configuration at C-2, C-3 and C-4.
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Word Map
- 1.1.3.10
- trametes
- multicolor
- 1,4-benzoquinone
- chrysosporium
- phanerochaete
-
white-rot
- nivale
-
microdochium
- l-sorbose
- aldopyranoses
- synthesis
- 1,5-anhydro-d-glucitol
-
flavinylated
-
ligninolytic
- ochracea
-
glucose-methanol-choline
- 1,5-anhydroglucitol
- peniophora
-
c4a-hydroperoxyflavin
- biotechnology
- food industry
- energy production
- biofuel production
- analysis
The taxonomic range for the selected organisms is: Trametes ochracea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
pyranose 2-oxidase, pyranose oxidase, carbohydrate oxidase, glucose 2-oxidase, pyranose-2-oxidase, pyranose/oxygen 2-oxidoreductase, tmp2ox, pyranose:oxygen-2-oxidoreductase, glucose-2-oxidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
P2O
-
pyranose 2-Oxidase
-
carbohydrate oxidase
-
-
-
-
glucose 2-oxidase
PROD
-
-
-
-
pyranose 2-Oxidase
-
-
pyranose-2-oxidase
-
-
-
-
glucose 2-oxidase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
highly regioselective mechanism, overview. The different conformations of the 454FSY456 gating segment in the semiopen and closed states induce backbone and side-chain movements of Thr169 and Asp452 that add further differential stabilization to the individual states
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
the enzyme shows a ping pong bi bi mechanism, a specialised bi bi mechanism in which substrate binding and release of products are ordered and the enzyme shuttles between a free and a substrate-modified intermediate conformation
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
investigation of the reaction mechanism by determining the rate constants
-
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
ping pong bi bi reaction mechanism, sugar oxidation and flavin reduction activation and mechanism, regiospecificity and selectivity of sugar oxidation, overview. The 2-oxo-sugar product is released prior to the oxygen reaction, overview. The enzyme shows a hydride transfer mechanism in which there is stepwise formation of D-glucose alkoxide prior to the hydride transfer, and a C4a-hydroperoxyflavin as an intermediate during the oxidative half-reaction, the C4a-hydroperoxyflavin merely eliminates H2O2 to generate oxidized FAD. The breakage of the flavin N (5)-H bond controls the overall process of H2O2 elimination from C4a-hydroperoxyflavin
-
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
ping pong bi bi reaction mechanism, the catalytic reaction of P2O can be divided into a reductive half-reaction in which two electrons are transferred as a hydride equivalent from a sugar substrate to a flavin to generate reduced FAD and a 2-oxo-sugar, and an oxidative half-reaction in which two electrons are transferred from the reduced flavin to oxygen to form hydrogen peroxide, formation of C(4a)-hydroperoxyflavin was observed during enzyme turnovers, overview
-
D-glucose + O2 = 2-dehydro-D-glucose + H2O2
ping-pong reaction mechanism, but change in the enzyme kinetic mechanism from a ping-pong type mechanism at pH values below 7.0 to a ternary complex type mechanism at pH values above 7.0. The switching of reaction mechanism from the ping-pong to the ternary complex type may be resulting from the change of structural dynamics of the substrate loop in the T169A mutant or at higher pH values, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
pyranose:oxygen 2-oxidoreductase
A flavoprotein (FAD). Also oxidizes D-xylose, L-sorbose and D-glucono-1,5-lactone, which have the same ring conformation and configuration at C-2, C-3 and C-4.
CAS REGISTRY NUMBER
COMMENTARY
37250-80-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-deoxy-2-fluoro-D-glucose + O2
2-deoxy-3-dehydro-D-glucose + H2O2
slow substrate
-
-
?
2-deoxy-D-galactose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
2-deoxy-D-glucose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
beta-D-galactose + O2
2-dehydro-D-galactose + H2O2
poor substrate
-
-
?
D-cellobiose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-fucose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-galactose + 1,4-benzoquinone
2-dehydro-D-galactose + hydroquinone
-
-
-
?
D-galactose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical
2-dehydro-D-galactose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)
-
-
-
?
D-galactose + ferricenium ion
2-dehydro-D-galactose + ferrocenium ion
-
-
-
r
D-galactose + ferrocenium hexafluorophosphate
2-dehydro-D-galactose + ferrocene
-
-
-
?
D-galactose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-glucose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical
2-dehydro-D-glucose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)
-
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
2-dehydro-D-glucose + reduced 2,6-dichlorophenolindophenol
D-glucose + ferrocenium hexafluorophosphate
2-dehydro-D-glucose + ferrocene
-
-
-
?
D-maltoheptaose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-maltopentaose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-maltotriose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-trehalose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-xylose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
L-arabinose + O2
?
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
5-thioglucose + O2
2-keto-5-thioglucose + H2O2
-
24% relative activity to D-glucose
-
?
D-galactose + 1,4-benzoquinone
2-dehydro-D-galactose + 1,4-hydroquinone
-
-
-
-
?
D-galactose + ferricenium hexafluorophosphate
2-dehydro-D-galactose + ferrocenium hexafluorophosphate
-
-
-
-
r
D-gluconolactone + O2
? + H2O2
-
64% relative activity to D-glucose
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + 1,4-hydroquinone
-
-
-
-
?
D-glucose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical
2-dehydro-D-glucose + 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)
-
-
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
2-dehydro-D-glucose + reduced 2,6-dichlorophenolindophenol
-
-
-
-
r
D-glucose + ferricenium hexafluorophosphate
2-dehydro-D-glucose + ferrocenium hexafluorophosphate
-
-
-
-
r
D-glucose + methyl-1,4-benzoquinone
2-dehydro-D-glucose + methylhydroquinone
-
-
-
-
r
D-glucose + tetrabromo-1,4-benzoquinone
2-dehydro-D-glucose + tetrabromohydroquinone
-
-
-
-
r
D-xylose + O2
D-xylosone + H2O2
-
56% relative activity to D-glucose
-
?
gentibiose + O2
2-keto-D-gentibiose + H2O2
-
23% relative activity to D-glucose
-
?
L-sorbose + O2
5-dehydro-D-fructose + H2O2
-
99% relative activity to D-glucose
-
?
D-galactose + 1,4-benzoquinone
2-dehydro-D-galactose + 1,4-hydroquinone
-
-
-
?
D-galactose + 1,4-benzoquinone
2-dehydro-D-galactose + 1,4-hydroquinone
-
-
-
-
?
D-galactose + 1,4-benzoquinone
2-dehydro-D-galactose + 1,4-hydroquinone
-
-
-
?
D-galactose + O2
2-dehydro-D-galactose + H2O2
His-tagged recombinant wild type enzyme strongly prefers D-glucose to D-galactose as its substrate
-
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + 1,4-hydroquinone
-
-
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + 1,4-hydroquinone
-
-
-
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + 1,4-hydroquinone
-
-
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + hydroquinone
-
-
-
?
D-glucose + 1,4-benzoquinone
2-dehydro-D-glucose + hydroquinone
1,4-benzoquinone is a physiologically relevant alternative electron acceptor in the oxidative half-reaction
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
2-dehydro-D-glucose + reduced 2,6-dichlorophenolindophenol
-
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
2-dehydro-D-glucose + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
D-glucose + ferricenium ion
2-dehydro-D-glucose + ferrocenium ion
-
-
-
?
D-glucose + ferricenium ion
2-dehydro-D-glucose + ferrocenium ion
-
-
-
-
?
D-glucose + ferricenium ion
2-dehydro-D-glucose + ferrocenium ion
-
-
-
r
D-glucose + O2
2-dehydro-D-glucose + H2O2
-
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
recombinant pyranose 2-oxidase analyzed
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
selectivity of pyranose 2-oxidase-based biosensor system for different sugar substrates analyzed
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
His-tagged recombinant wild type enzyme strongly prefers D-glucose to D-galactose as its substrate
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
the wild type enzyme displays a clear preference for D-glucose over D-galactose
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
highly regioselective mechanism
-
-
?
D-galactose + O2
2-dehydro-D-galactose + H2O2
-
5.7% relative activity to D-glucose
-
?
D-galactose + O2
2-dehydro-D-galactose + H2O2
-
mutants T169S, T169N, and T169G
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
-
-
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
-
immobilized and soluble pyranose 2-oxidase analyzed
-
-
?
D-glucose + O2
D-glucosone + H2O2
-
D-glucose is the preferred substrate
-
?
additional information
?
-
ferulic acid oxidation in a model system and of thiol oxidation in a wheat flour extract, overview. L-arabinose is a poor substrate. Dehydroascorbic acid does not act as electron acceptor
-
-
?
additional information
?
-
-
pyranose 2-oxidase from Trametes multicolor is a flavoenzyme that catalyzes the oxidation of D-glucose and other aldopyranose sugars at the C2 position by using O2 as an electron acceptor to form the corresponding 2-oxo-sugars and H2O2
-
-
?
additional information
?
-
-
most of the catalytic dehydrogenation of substrates by flavoprotein oxidases in the GMC class is initiated by the removal of a hydroxyl proton followed by the transfer of a hydride moiety. Pyranose 2-oxidase catalyzes the oxidation of several aldopyranoses by molecular oxygen at the C2 position to yield the corresponding 2-keto-aldoses and hydrogen peroxide
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-galactose + O2
2-dehydro-D-galactose + H2O2
-
mutants T169S, T169N, and T169G
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
-
-
-
-
?
additional information
?
-
-
pyranose 2-oxidase from Trametes multicolor is a flavoenzyme that catalyzes the oxidation of D-glucose and other aldopyranose sugars at the C2 position by using O2 as an electron acceptor to form the corresponding 2-oxo-sugars and H2O2
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
-
-
-
?
D-glucose + O2
2-dehydro-D-glucose + H2O2
highly regioselective mechanism
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
the type of aminoacyl flavin covalent link is 8alpha-(N3-histidyl)-FAD
FAD
-
each subunit contains one FAD covalently linked at the C8 methyl group of FAD to Ne2 of residue His167
FAD
-
one FAD per subunit, Trp168 residue near the flavin isoalloxazine ring can potentially act as an electron donor to the excited flavin isoalloxazine ring, isoalloxazine is covalently linked to His167
FAD
-
the FAD cofactor in P2O is covalently linked through a histidyl linkage at His167
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-keto-D-glucose
-
5% reduction at 100 mM, 10% reduction at 200 mM, no deactivating effect determined
H2O2
-
oxidation of Phe454, which is located at the flexible active-site loop of the enzyme, is the first and main step in the inactivation of the enzyme by hydrogen peroxide
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.07
2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) cation radical
-
pH 6.5, substrate: D-glucose
0.0052
1,4-benzoquinone
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0071
1,4-benzoquinone
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0089
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.01
1,4-benzoquinone
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.013
1,4-benzoquinone
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.027
1,4-benzoquinone
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.029
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.029
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.032
1,4-benzoquinone
L537W/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.036
1,4-benzoquinone
L537W mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.037
1,4-benzoquinone
L537G/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.038
1,4-benzoquinone
L537W/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.04
1,4-benzoquinone
E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.04
1,4-benzoquinone
L537G/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.048
1,4-benzoquinone
L537G mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.049
1,4-benzoquinone
E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.052
1,4-benzoquinone
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.065
1,4-benzoquinone
wild-type, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.072
1,4-benzoquinone
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.072
1,4-benzoquinone
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.078
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.13
1,4-benzoquinone
L537W mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.136
1,4-benzoquinone
E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.137
1,4-benzoquinone
L537W/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.14
1,4-benzoquinone
L537W/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.14
1,4-benzoquinone
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.15
1,4-benzoquinone
L537G/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.155
1,4-benzoquinone
L537G/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.176
1,4-benzoquinone
L537G mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.182
1,4-benzoquinone
E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.24
1,4-benzoquinone
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.241
1,4-benzoquinone
wild-type, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.093
D-galactose
T169G/E542K/V546C, mutant, 30°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.19
D-galactose
T169G/E542K/V546C, mutant, 50°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.23
D-galactose
wild-type, mutant, 50°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.25
D-galactose
wild-type, mutant, 30°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.4
D-galactose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
0.983
D-galactose
V546P/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.66
D-galactose
T169G/E542K/V546C, mutant, 30°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
2.45
D-galactose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
2.45
D-galactose
mutant enzyme H450G, pH and temperature not specified in the publication
2.48
D-galactose
T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.48
D-galactose
mutant enzyme T169G, pH and temperature not specified in the publication
2.76
D-galactose
T169G/E542K/V546C, mutant, 50°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
3.56
D-galactose
T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
3.87
D-galactose
E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
3.87
D-galactose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
4.26
D-galactose
E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.19
D-galactose
L537W/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.49
D-galactose
L537W/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.77
D-galactose
L537G/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
6.01
D-galactose
L537G/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
6.1
D-galactose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
6.38
D-galactose
R472G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
7.21
D-galactose
R472L, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
7.39
D-galactose
N593R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
7.8
D-galactose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
7.94
D-galactose
wild-type, mutant, 30°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
8.09
D-galactose
wild-type, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
8.79
D-galactose
wild-type, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
8.79
D-galactose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
8.79
D-galactose
wild type enzyme, pH and temperature not specified in the publication
9.27
D-galactose
H548R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
9.4
D-galactose
L537W mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
9.47
D-galactose
L537G mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
9.89
D-galactose
H548I, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
10
D-galactose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
10.4
D-galactose
V546C/T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
12
D-galactose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
12
D-galactose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
12.9
D-galactose
T169S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
13
D-galactose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
13
D-galactose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
14.6
D-galactose
wild-type, mutant, 50°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
26
D-galactose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
29
D-galactose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
29.4
D-galactose
V546G/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
34
D-galactose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
46.2
D-galactose
V546C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
46.2
D-galactose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
46.2
D-galactose
mutant enzyme V546C, pH and temperature not specified in the publication
50.6
D-galactose
V546P, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
200
D-galactose
V546G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
750
D-galactose
Q448S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
940
D-galactose
Q448N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1260
D-galactose
Q448C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.018
D-glucose
mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.042
D-glucose
mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.046
D-glucose
mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C
0.057
D-glucose
mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.082
D-glucose
mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C
0.092
D-glucose
mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C
0.12
D-glucose
mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
0.13
D-glucose
wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C
0.14
D-glucose
mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.14
D-glucose
mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.18
D-glucose
mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
0.22
D-glucose
T169G/E542K/V546C, mutant, 30°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.23
D-glucose
mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
0.24
D-glucose
mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
0.24
D-glucose
wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.25
D-glucose
wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
0.31
D-glucose
T169G/E542K/V546C, mutant, 50°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.32
D-glucose
wild-type protein, 0.1 M ethanolamine buffer at pH 10.5
0.327
D-glucose
T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.35
D-glucose
recombinant protein with a hexa-histidine tag, 0.1 M ethanolamine buffer at pH 10.5
0.394
D-glucose
T169S, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.4
D-glucose
wild-type, mutant, 30°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.4
D-glucose
mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C
0.419
D-glucose
L537W/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.432
D-glucose
L537W/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.44
D-glucose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
0.441
D-glucose
L537G/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.487
D-glucose
L537G/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.489
D-glucose
E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.521
D-glucose
E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.521
D-glucose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.527
D-glucose
N593R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.56
D-glucose
mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
0.64
D-glucose
T169G/E542K/V546C, mutant, 30°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.658
D-glucose
V546P/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.69
D-glucose
mutant enzyme T169G, pH and temperature not specified in the publication
0.691
D-glucose
T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.698
D-glucose
wild-type, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.72
D-glucose
mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
0.749
D-glucose
L537W mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.76
D-glucose
wild-type, mutant, 30°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.76
D-glucose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.77
D-glucose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.773
D-glucose
R472G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.78
D-glucose
wild-type, mutant, 50°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.8 - 11
D-glucose
H548I, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.81
D-glucose
mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
0.851
D-glucose
L537G mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.88
D-glucose
H548R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.886
D-glucose
R472L, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.939
D-glucose
wild-type, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.939
D-glucose
wild type enzyme, pH and temperature not specified in the publication
0.952
D-glucose
V546C/T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.98
D-glucose
mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C
0.984
D-glucose
V546G/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.987
D-glucose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.987
D-glucose
mutant enzyme H450G, pH and temperature not specified in the publication
0.99
D-glucose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
1.15
D-glucose
T169G/E542K/V546C, mutant, 50°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
1.18
D-glucose
wild-type, mutant, 50°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
1.5
D-glucose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.5
D-glucose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.51
D-glucose
E542K with a hexa-histidine tag, 0.1 M ethanolamine buffer at pH 10.5
1.6
D-glucose
mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
1.7
D-glucose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.7
D-glucose
mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
1.8
D-glucose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.9
D-glucose
wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
2.1
D-glucose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2.1
D-glucose
mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
2.2
D-glucose
V546P, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.43
D-glucose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
2.43
D-glucose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
2.5
D-glucose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3.06
D-glucose
V546C, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
3.06
D-glucose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
3.06
D-glucose
mutant enzyme V546C, pH and temperature not specified in the publication
5.7
D-glucose
V546G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
5.97
D-glucose
mutant enzyme H167A, pH and temperature not specified in the publication
28
D-glucose
Q448N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
30
D-glucose
Q448S, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
100
D-glucose
Q448C, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2 - 3
D-melibiose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
50
D-melibiose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
240
D-melibiose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
260
D-melibiose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
350
D-melibiose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
390
D-melibiose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1500
D-melibiose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.054
ferricenium ion
L537G/E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.063
ferricenium ion
L537W mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.068
ferricenium ion
E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.07
ferricenium ion
wild-type, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.072
ferricenium ion
L537G/E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.074
ferricenium ion
L537W/E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.086
ferricenium ion
L537G mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.09
ferricenium ion
L537W/E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.183
ferricenium ion
E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.253
ferricenium ion
L537W mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.254
ferricenium ion
wild-type, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.281
ferricenium ion
L537W/E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.289
ferricenium ion
L537G mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.29
ferricenium ion
E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.296
ferricenium ion
L537G/E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.319
ferricenium ion
E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.328
ferricenium ion
L537G/E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.408
ferricenium ion
L537W/E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
0.015
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.016
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.023
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.025
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.041
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.049
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.1
ferrocenium hexafluorophosphate
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.14
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.15
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.24
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.28
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.35
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.38
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.4
ferrocenium hexafluorophosphate
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
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
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
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
0.3
1,4-benzoquinone
-
soluble enzyme, 500 mM glucose used as electron acceptor, pH 5.0
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
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
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
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
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
3.1
D-galactose
-
mutant enzyme T169G, in 50 mM sodium phosphate (pH 7.0), at 25°C
15
D-galactose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 25°C
16
D-galactose
-
mutant enzyme T169N, in 50 mM sodium phosphate (pH 7.0), at 25°C
16.2
D-galactose
-
mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 25°C
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
50
D-galactose
-
mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 25°C
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
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
0.9
D-glucose
-
mutant enzyme T169G, in 50 mM sodium phosphate (pH 7.0), at 4°C
1.13
D-glucose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 4°C
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
1.7
D-glucose
-
mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 4°C
1.9
D-glucose
-
O2 concentration constant, pH 7.0, absorbance at 420 nm resulting from the oxidation of diammonium-2,2'-azinobis-(3-ethylbenzthiazolin-6-sulfonic acid) by H2O2
2.3
D-glucose
-
wild type enzyme, at pH 6.5, temperature not specified in the publication
2.5
D-glucose
-
mutant enzyme N593C, at pH 6.5, temperature not specified in the publication
3.2
D-glucose
-
O2 concentration constant, pH 7.0, calculated with kinetic equation
4.6
D-glucose
-
mutant enzyme T169N, in 50 mM sodium phosphate (pH 7.0), at 4°C
30
D-glucose
-
mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 4°C
45
D-glucose
-
pH 7.0, 25°C, reductive half-reaction, wild-type enzyme and mutant T169S
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
0.092
ferricenium hexafluorophosphate
-
mutant enzyme V546C/T169G/L537W, at pH 6.5 and 30°C
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
0.15
ferricenium hexafluorophosphate
-
mutant enzyme V546C/E542K, at pH 6.5 and 30°C
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
0.22
ferricenium hexafluorophosphate
-
mutant enzyme V546C/T169G/L537W, at pH 6.5 and 30°C
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
0.31
ferricenium hexafluorophosphate
-
mutant enzyme V546C/T169G, at pH 6.5 and 30°C
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
0.5
ferricenium hexafluorophosphate
-
mutant enzyme V546C/T169G, at pH 6.5 and 30°C
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
0.79
glucose
-
immobilized enzyme, O2 used as electron acceptor, determined in cuvette assay, pH 5.0
0.85
glucose
-
soluble enzyme, O2 used as electron acceptor, determined in cuvette assay, pH 5.0
0.03
O2
-
wild type enzyme, using D-galactose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.04
O2
-
pH 5.5, substrate oxygen limited (D-glucose excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.04
O2
-
pH 8.5, substrate D-glucose limited (oxygen excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.07
O2
-
mutant enzyme T169S, using D-galactose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.09
O2
-
wild type enzyme, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.1
O2
-
pH 6.5, substrate oxygen limited (D-glucose excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.1
O2
-
pH 7.5, substrate oxygen limited (D-glucose excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.132
O2
-
glucose concentration constant, pH 7.0, calculated with kinetic equation
0.2
O2
-
pH 7.5, substrate D-glucose limited (oxygen excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.2
O2
-
mutant enzyme T169A, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.22
O2
-
D-glucose concentration constant, pH 7.0, absorbance at 420 nm resulting from the oxidation of diammonium-2,2'-azinobis-(3-ethylbenzthiazolin-6-sulfonic acid) by H2O2
0.3
O2
-
pH 5.5, substrate D-glucose limited (oxygen excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.4
O2
-
mutant enzyme T169A, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
0.5
O2
-
pH 8.5, substrate oxygen limited (D-glucose excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.9
O2
-
pH 6.5, substrate D-glucose limited (oxygen excess), measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
0.99
O2
-
mutant enzyme T169S, using D-glucose as cosubstrate ,in 50 mM sodium phosphate (pH 7.0), at 4°C
1.32
O2
-
mutant enzyme T169N, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
3.7
O2
-
mutant enzyme T169G, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
4.5
O2
-
mutant enzyme T169G, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
4.8
O2
-
mutant enzyme T169N, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
additional information
additional information
Michaelis-Menten kinetics, overview
-
additional information
additional information
D-mannose, glucosamine, beta-lactose, sucrose and D-maltose lack response signal, higher maltooligosaccharides good substrates for pyranose 2-oxidase-based biosensor
-
additional information
additional information
-
kinetic parameters of the reaction measured (substrates D-glucose or 2-deoxy-D-glucose), pH 7.0
-
additional information
additional information
-
steady-state and pre-steady-state kinetics, and kinetic isotope effects, of oxidative half-reaction and overall reaction, overview
-
additional information
additional information
-
the steady-state kinetics of P2O can be classified as a ping pong bi-bi type, because the 2-keto-sugar product is released prior to the oxygen reaction, transient kinetics and isotope effects, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
324
1,4-benzoquinone
-
soluble enzyme, 500 mM glucose used as electron acceptor, pH 5.0
49
glucose
-
soluble enzyme, O2 used as electron acceptor, determined in cuvette assay, pH 5.0
1.2
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2
1,4-benzoquinone
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2.7
1,4-benzoquinone
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2.9
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3
1,4-benzoquinone
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3.3
1,4-benzoquinone
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3.8
1,4-benzoquinone
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
4.37
1,4-benzoquinone
E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
4.64
1,4-benzoquinone
L537G mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
4.72
1,4-benzoquinone
L537G/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
4.75
1,4-benzoquinone
L537G/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
4.77
1,4-benzoquinone
L537W/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
4.79
1,4-benzoquinone
wild-type, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.09
1,4-benzoquinone
L537W/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.37
1,4-benzoquinone
L537W mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.52
1,4-benzoquinone
E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
15
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
30
1,4-benzoquinone
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
61
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
127
1,4-benzoquinone
E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
130
1,4-benzoquinone
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
152
1,4-benzoquinone
wild-type, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
160
1,4-benzoquinone
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
173
1,4-benzoquinone
L537G/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
173
1,4-benzoquinone
L537G/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
175
1,4-benzoquinone
L537W/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
181
1,4-benzoquinone
L537W/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
184
1,4-benzoquinone
L537G mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
189
1,4-benzoquinone
E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
205
1,4-benzoquinone
L537W mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
220
1,4-benzoquinone
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
220
1,4-benzoquinone
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.14
D-galactose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.245
D-galactose
V546P/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.27
D-galactose
T169G/E542K/V546C, mutant, 30°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.27
D-galactose
mutant enzyme T169G, pH and temperature not specified in the publication
0.273
D-galactose
T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.3
D-galactose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.331
D-galactose
V546G/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.356
D-galactose
H548R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.38
D-galactose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
0.433
D-galactose
Q448N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.622
D-galactose
Q448C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.64
D-galactose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.654
D-galactose
V546C/T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.74
D-galactose
T169G/E542K/V546C, mutant, 50°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
1.2
D-galactose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.5
D-galactose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.53
D-galactose
Q448S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.57
D-galactose
N593R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.6
D-galactose
V546P, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.69
D-galactose
R472L, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.7
D-galactose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.96
D-galactose
T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
1.99
D-galactose
E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2
D-galactose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2.07
D-galactose
R472G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.1
D-galactose
wild-type, mutant, 30°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
2.34
D-galactose
L537G/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.36
D-galactose
L537G/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.48
D-galactose
L537W/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.51
D-galactose
L537W/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.51
D-galactose
wild-type, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.51
D-galactose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
2.51
D-galactose
wild type enzyme, pH and temperature not specified in the publication
2.56
D-galactose
L537G mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.59
D-galactose
E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.59
D-galactose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
2.66
D-galactose
H548I, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.73
D-galactose
wild-type, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.84
D-galactose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
2.9
D-galactose
L537W mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
3.51
D-galactose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
3.51
D-galactose
mutant enzyme H450G, pH and temperature not specified in the publication
4.51
D-galactose
V546G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
5.51
D-galactose
wild-type, mutant, 50°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
5.53
D-galactose
T169S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
5.92
D-galactose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
5.92
D-galactose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
6.57
D-galactose
V546C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
6.57
D-galactose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
6.57
D-galactose
mutant enzyme V546C, pH and temperature not specified in the publication
6.61
D-galactose
wild-type, mutant, 30°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
14.6
D-galactose
wild-type, mutant, 50°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
171
D-galactose
T169G/E542K/V546C, mutant, 50°C, substrate D-galactose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.072
D-glucose
T169G/E542K/V546C, mutant, 30°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.119
D-glucose
V546G/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.18
D-glucose
mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C
0.2
D-glucose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.232
D-glucose
V546P/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.26
D-glucose
mutant enzyme T169G, pH and temperature not specified in the publication
0.262
D-glucose
T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.35
D-glucose
T169G/E542K/V546C, mutant, 50°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.38
D-glucose
mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C
0.43
D-glucose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
0.963
D-glucose
V546C/T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
0.99
D-glucose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.1
D-glucose
mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C
1.51
D-glucose
Q448C, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.06
D-glucose
mutant enzyme H167A, pH and temperature not specified in the publication
2.41
D-glucose
T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
2.7
D-glucose
mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C
3.9
D-glucose
mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
4.6
D-glucose
mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
4.73
D-glucose
Q448N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
5.42
D-glucose
Q448S, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
6.81
D-glucose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
7.1
D-glucose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
8.1
D-glucose
mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
9.2
D-glucose
H548R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
11
D-glucose
mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C
12
D-glucose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
12
D-glucose
mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
12.5
D-glucose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
12.5
D-glucose
mutant enzyme H450G, pH and temperature not specified in the publication
15.6
D-glucose
N593R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
15.8
D-glucose
V546P, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
16.8
D-glucose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
16.8
D-glucose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
17
D-glucose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
21.16
D-glucose
T169G/E542K/V546C, mutant, 30°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
21.8
D-glucose
T169S, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
22
D-glucose
mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
23.6
D-glucose
V546G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
26
D-glucose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
26
D-glucose
mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
27
D-glucose
mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
28.5
D-glucose
E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
30.9
D-glucose
R472L, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
31.6
D-glucose
H548I, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
31.7
D-glucose
L537W/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
32.2
D-glucose
R472G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
33
D-glucose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
33.1
D-glucose
L537G/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
34
D-glucose
wild-type, mutant, 30°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
35.4
D-glucose
wild-type, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
35.9
D-glucose
E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
35.9
D-glucose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
42
D-glucose
wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C
43.9
D-glucose
L537G/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
46.5
D-glucose
L537W/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
48.1
D-glucose
wild-type, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
48.1
D-glucose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
48.1
D-glucose
wild type enzyme, pH and temperature not specified in the publication
50
D-glucose
mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
52.1
D-glucose
L537G mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
58.9
D-glucose
wild-type, mutant, 50°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
59
D-glucose
L537W mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
70
D-glucose
wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
79.3
D-glucose
T169G/E542K/V546C, mutant, 50°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
88.6
D-glucose
V546C, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay
88.6
D-glucose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
88.6
D-glucose
mutant enzyme V546C, pH and temperature not specified in the publication
110
D-glucose
mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
134
D-glucose
mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
134
D-glucose
mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
142
D-glucose
mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
151
D-glucose
wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
152
D-glucose
wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
166
D-glucose
mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
265
D-glucose
mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
349
D-glucose
wild-type, mutant, 30°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
442
D-glucose
mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
615
D-glucose
wild-type, mutant, 50°C, substrate D-glucose (1,4-benzoquinone saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay
0.19
D-melibiose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.29
D-melibiose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.3
D-melibiose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2.7
D-melibiose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3.3
D-melibiose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
4.4
D-melibiose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
7.6
D-melibiose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1.44
ferricenium ion
E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
1.81
ferricenium ion
L537G/E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.08
ferricenium ion
E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.11
ferricenium ion
L537G/E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.47
ferricenium ion
L537W/E542R mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.68
ferricenium ion
L537W/E542K mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
5.34
ferricenium ion
wild-type, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
7.07
ferricenium ion
L537G mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
8.18
ferricenium ion
L537W mutant, substrate ferricenium ion (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
46.7
ferricenium ion
E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
54.4
ferricenium ion
E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
86.3
ferricenium ion
L537W/E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
86.7
ferricenium ion
L537G/E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
102
ferricenium ion
L537G/E542R mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
127
ferricenium ion
L537W/E542K mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
151
ferricenium ion
wild-type, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
282
ferricenium ion
L537G mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
334
ferricenium ion
L537W mutant, substrate ferricenium ion (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C
2.9
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
4.5
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
5.4
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
6.6
ferrocenium hexafluorophosphate
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
7.3
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
7.3
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
17
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
67
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
74
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
110
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
210
ferrocenium hexafluorophosphate
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
400
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
420
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
470
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.7
D-galactose
-
mutant enzyme T169G, in 50 mM sodium phosphate (pH 7.0), at 25°C
1.2
D-galactose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 25°C
13.7
D-galactose
-
mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.06
D-glucose
-
mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 4°C
5.6
D-glucose
-
mutant enzyme T169N, in 50 mM sodium phosphate (pH 7.0), at 4°C
8.15
D-glucose
-
pH 7.0, absorbance at 420 nm resulting from the oxidation of diammonium-2,2'-azinobis-(3-ethylbenzthiazolin-6-sulfonic acid) by H2O2
9.7
D-glucose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 4°C
5.9
O2
-
pH 5.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
6.5
O2
-
pH 7.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
6.8
O2
-
pH 6.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
7.5
O2
-
pH 8.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
37
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
130
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
140
1,4-benzoquinone
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
230
1,4-benzoquinone
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
280
1,4-benzoquinone
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
330
1,4-benzoquinone
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
420
1,4-benzoquinone
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
520
1,4-benzoquinone
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
520
1,4-benzoquinone
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
940
1,4-benzoquinone
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1200
1,4-benzoquinone
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2100
1,4-benzoquinone
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
2600
1,4-benzoquinone
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
3000
1,4-benzoquinone
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.011
D-galactose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.024
D-galactose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.046
D-galactose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.053
D-galactose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.059
D-galactose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.062
D-galactose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.11
D-galactose
mutant enzyme T169G, pH and temperature not specified in the publication
0.142
D-galactose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.142
D-galactose
mutant enzyme V546C, pH and temperature not specified in the publication
0.27
D-galactose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.286
D-galactose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.286
D-galactose
wild type enzyme, pH and temperature not specified in the publication
0.364
D-galactose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.493
D-galactose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.493
D-galactose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
0.669
D-galactose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
0.94
D-galactose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
1.43
D-galactose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
1.43
D-galactose
mutant enzyme H450G, pH and temperature not specified in the publication
0.094
D-glucose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.18
D-glucose
mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C
0.35
D-glucose
mutant enzyme H167A, pH and temperature not specified in the publication
0.38
D-glucose
mutant enzyme T169G, pH and temperature not specified in the publication
0.54
D-glucose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.99
D-glucose
mutant enzyme T196G/V546C, pH and temperature not specified in the publication
2.8
D-glucose
mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C
4.6
D-glucose
mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C
4.7
D-glucose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
7
D-glucose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
8.2
D-glucose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
8.83
D-glucose
mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
12.7
D-glucose
mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C
12.7
D-glucose
mutant enzyme H450G, pH and temperature not specified in the publication
13.5
D-glucose
mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
13.5
D-glucose
mutant enzyme H450G/V546C, pH and temperature not specified in the publication
15
D-glucose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
22
D-glucose
mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
27
D-glucose
mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
29
D-glucose
mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C
29
D-glucose
mutant enzyme V546C, pH and temperature not specified in the publication
32
D-glucose
mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
34
D-glucose
mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
38
D-glucose
wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
43
D-glucose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
48
D-glucose
mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
51.2
D-glucose
His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C
51.2
D-glucose
wild type enzyme, pH and temperature not specified in the publication
58
D-glucose
mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C
68.9
D-glucose
mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C
70
D-glucose
mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
79
D-glucose
mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
120
D-glucose
mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C
127
D-glucose
mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C
203
D-glucose
mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
281
D-glucose
mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
308
D-glucose
wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C
584
D-glucose
mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
592
D-glucose
wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
632
D-glucose
wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
1193
D-glucose
mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C
1213
D-glucose
mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
1465
D-glucose
mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
2608
D-glucose
mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C
0.0037
D-melibiose
mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.005
D-melibiose
wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0058
D-melibiose
mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.008
D-melibiose
mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0084
D-melibiose
mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0113
D-melibiose
mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.0166
D-melibiose
mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
63
ferrocenium hexafluorophosphate
wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
180
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
180
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
200
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
290
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
340
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
360
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
370
ferrocenium hexafluorophosphate
mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
480
ferrocenium hexafluorophosphate
mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
500
ferrocenium hexafluorophosphate
mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
510
ferrocenium hexafluorophosphate
wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1210
ferrocenium hexafluorophosphate
mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1250
ferrocenium hexafluorophosphate
mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
1640
ferrocenium hexafluorophosphate
mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C
0.005
D-galactose
-
mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.08
D-galactose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.15
D-galactose
-
mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.3
D-galactose
-
mutant enzyme T169N, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.5
D-galactose
-
mutant enzyme T169G, in 50 mM sodium phosphate (pH 7.0), at 25°C
0.002
D-glucose
-
mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.8
D-glucose
-
mutant enzyme T169G, in 50 mM sodium phosphate (pH 7.0), at 4°C
1.2
D-glucose
-
mutant enzyme T169N, in 50 mM sodium phosphate (pH 7.0), at 4°C
8.1
D-glucose
-
mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 4°C
8.6
D-glucose
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.07
O2
-
mutant enzyme T169A, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
0.2
O2
-
mutant enzyme T169G, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.4
O2
-
mutant enzyme T169A, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
0.4
O2
-
mutant enzyme T169G, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
0.99
O2
-
mutant enzyme T169S, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
3.4
O2
-
mutant enzyme T169N, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
4.8
O2
-
mutant enzyme T169N, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
32
O2
-
mutant enzyme T169S, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C
44.2
O2
-
wild type enzyme, using D-galactose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 25°C
110
O2
-
wild type enzyme, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
82.2
additional information
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
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
additional information
comparative characterization of pyranose 2-oxidase from Trametes ochracea and Trametes pubescens, enzyme activity determined by the formation of H2O2 in a peroxidase-coupled assay, inclusion body formation and specific activity of pyranose 2-oxidases in Escherichia coli harbouring chimeric plasmid constructs, site-directed recombination
additional information
-
comparative characterization of pyranose 2-oxidase from Trametes ochracea and Trametes pubescens, enzyme activity determined by the formation of H2O2 in a peroxidase-coupled assay, inclusion body formation and specific activity of pyranose 2-oxidases in Escherichia coli harbouring chimeric plasmid constructs, site-directed recombination
additional information
pyranose 2-oxidase-based biosensor system, electrical wiring of different types of pyranose oxidase with an osmium redox polymer on graphite electrodes, optimization studies using glucose as substrate, biosensors with best characteristics in terms of linear range, detection limit and sensitivity used for analyzing selectivity for different sugar substrates
additional information
kinetic parameters measured at batch conversion experiments of wild-type pyranose oxidase and mutant T169G/E542K/V546C
additional information
-
kinetic parameters measured at batch conversion experiments of wild-type pyranose oxidase and mutant T169G/E542K/V546C
additional information
-
report on immobilization of pyranose 2-oxidase on agarose or acrylic resins using different coupling methods, enzyme activity determined by a peroxidase-coupled assay or by measuring initial rate of oxygen consumption, molecular properties of immobilized enzymes and binding capacities summarized
additional information
-
complete oxidation of D-glucose using pyranose oxidase as the synthesizing biocatalyst and laccase for redox mediator regeneration is described quantitatively to increase both stability and productivity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8 - 11
optimal buffer combination for the pyranose 2-oxidase systems determined
5.5 - 8.5
-
substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C, varying concentrations of both D-glucose and oxygen. Ping-pong kinetic mechanism at pH values below 7.0 and an ordered mechanism at pH above 7.0
additional information
-
flavin oxidation occurs via different pathways depending on the pH of the environment. At pH values lower than 8.0, the reduced enzyme reacts with O2 to form a C4a-hydroperoxyflavin intermediate, leading to elimination of H2O2. At pH 8.0 and higher, the majority of the reduced enzyme reacts with O2 via a pathway that does not allow detection of the C4a-hydroperoxyflavin, and flavin oxidation occurs with decreased rate constants upon the rise in pH. The switching between the two modes of enzyme oxidation is controlled by protonation of a group which has a pKa of 7.6, stopped-flow spectrophotometry, overview. The protonation of the group which controls the mode of flavin oxidation cannot be rapidly equilibrated with outside solvent. Using a double-mixing stopped-flow experiment, a rate constant for proton dissociation from the reaction site is determined to be 21.0 s-1
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
additional information
evolution
-
the enzyme belongs to the glucose-methanol-choline (GMC) oxidoreductase superfamily
evolution
-
the enzyme belongs to the glucose-methanol-choline (GMC) oxidoreductase superfamily, comparison of P2O and other enzymes in the GMC family, overview. Although all of the GMC enzymes share similar structural folding and use the hydride transfer mechanism for flavin reduction, they appear to have subtle differences in the fine-tuned details of how they catalyze substrate oxidation
additional information
substrate recognition loop, active-site protein interactions for D-glucose, and structural determinants of the productive glucose C2-oxidation mode, detailed overview
additional information
-
substrate recognition loop, active-site protein interactions for D-glucose, and structural determinants of the productive glucose C2-oxidation mode, detailed overview
additional information
-
comparison the P2O active site in H167A, T169S or T169S-acetate complex with wild-type P2O-acetate complex, molecuar dynamics and spectroscopic analysi, detailed overview
additional information
-
flavin oxidation occurs via different pathways depending on the pH of the environment. At pH values lower than 8.0, the reduced enzyme reacts with O2 to form a C4a-hydroperoxyflavin intermediate, leading to elimination of H2O2. At pH 8.0 and higher, the majority of the reduced enzyme reacts with O2 via a pathway that does not allow detection of the C4a-hydroperoxyflavin, and flavin oxidation occurs with decreased rate constants upon the rise in pH. The switching between the two modes of enzyme oxidation is controlled by protonation of a group which has a pKa of 7.6, stopped-flow spectrophotometry, overview
additional information
-
redox potential of wild-type and mutant enzymes, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). Q7ZA32_TRAOC
623
0
69342
TrEMBL
Mitochondrion (Reliability: 4)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
270000
270000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
-
4 * 65000, SDS-PAGE, corroborated by electron microscopy studies
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 0.1 M MES (pH 5.2), 50 mM MgCl2, and 10% (w/v) monomethylether polyethylene glycol 2000
mutant enzymes are crystallized by the hanging drop vapor diffusion method, using 10% (w/v) monomethyl ether poly(ethlene glycol) 2000 , 0.1 M MES (pH 5.2), 50 mM MgCl2
mutant enzymes H167A and T169, Ahanging drop vapor diffusion method, using 0.1 M sodium acetate (pH 4.6), 50 mM MgCl2, and 6% (w/v) monomethyl ether PEG 2000
P2O H167A mutant in complex with 3-deoxy-3-fluoro-D-glucose, microseeded hanging drops, mixing of equal volumes of 0.1 M MES, pH 5.2, 50 mM MgCl2, 10% w/v PEG monomethyl ether 2000 with 20 mg/ml protein in 20 mM MES, pH 5.2, addition of ligand in saturated mother liquor with 28% w/v PEG 2000, X-ray diffraction structure determination and analysis at 1.35 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D452A/R472A
the mutant shows drastic effects on the binding constant for D-glucose
E542K
F454A/S455A/Y456A
the mutant shows decreased catalytic efficiency for D-glucose/O2 compared to the wild type enzyme
F454A/Y456A
the mutant shows decreased catalytic efficiency for D-glucose/O2 compared to the wild type enzyme
F454N
F454P
the mutant shows strongly decreased catalytic efficiency for D-glucose/O2 compared to the wild type enzyme
H167A
H450G
H450G/E542K/V546C
mutant shows catalytic efficiency for its substrate D-galactose that is increased 1.2fold compared to the wild type enzyme while kcat/KM for D-glucose is decreased to 17% of its original value
H450G/V546C
H450Q
the turnover number for the electron donor/acceptor substrate pair D-galactose/O2 is comparable to that of the wild type enzyme
L545C
L547R
Q448H
T166R
T169G
T169G/E542K/V546C
mutant, analysis of kinetic parameters and thermal stability
T169N
T169S
T196G/V546C
the mutant exhibits reduced D-glucose conversion and D-galactose conversion activities
V546C
Y456W
the turnover number for the electron donor/acceptor substrate pair D-galactose/O2 is comparable to that of the wild type enzyme
F454A
-
the mutant shows about 40fold reduced catalytic efficiency compared to the wild type enzyme
F454Y
-
the mutant shows about 3.5fold reduced catalytic efficiency compared to the wild type enzyme
H167A
H167A/H548A
-
site-directed mutagenesis, reductively inactive mutant, contains noncovalently linked FAD
H167A/H548D
-
site-directed mutagenesis, reductively inactive mutant, contains noncovalently linked FAD
H167A/H548N
-
site-directed mutagenesis, reductively inactive mutant, contains noncovalently linked FAD
H167A/H548R
H167A/H548S
-
site-directed mutagenesis, reductively inactive mutant, contains noncovalently linked FAD
H169S
-
site-directed mutagenesis, spectroscopic analysis and molecular dynamics of the T169S P2O-acetate complex
H548A
-
site-directed mutagenesis, reductive activity of the mutant is reduced compared to the wild-type enzyme, contains noncovalently and covalently linked FAD
H548D
-
site-directed mutagenesis, reductive activity of the mutant is reduced compared to the wild-type enzyme, contains noncovalently and covalently linked FAD
H548N
-
site-directed mutagenesis, reductive activity of the mutant is reduced compared to the wild-type enzyme, contains noncovalently and covalently linked FAD
H548R
-
site-directed mutagenesis, reductive activity of the mutant is slightly reduced compared to the wild-type enzyme, contains some noncovalently and mostly covalently linked FAD
H548S
-
site-directed mutagenesis, reductive activity of the mutant is reduced compared to the wild-type enzyme, contains noncovalently and covalently linked FAD
N593C
-
the mutant shows a complete loss of the oxidase function with full preservation of substrate (dehydrogenase) activity
N593H
-
site-directed mutagenesis, the enzyme contains a covalently linked FAD, similar to the wild-type enzyme
T169A
T169G
T169N
T169S
additional information
E542K
the mutant is characterized by reduced KM values for both D-glucose and D-galactose and significantly increased stability
F454N
the apparent kcat value with D-glucose and O2 (fixed at a concentration of 0.256 mM, air saturation) as substrates decreases dramatically compared to the wild type enzyme
F454N
the mutant shows decreased catalytic efficiency for D-glucose/O2 compared to the wild type enzyme
H167A
the P2O mutant in complex with the slow substrate 2-deoxy-2-fluoro-D-glucose shows a substrate-binding mode compatible with oxidation at C3. To accommodate the sugar, a gating segment, 454FSY456, in the substrate recognition loop partly unfolded to create a spacious and more polar active site that is distinct from the closed state of P2O. With 3-deoxy-3-fluoro-D-glucose the mutant prefers C2 oxidation, crystal structure of the ordered complex of P2O H167A with at 1.35 A resolution, overview
H167A
the mutant exhibits reduced D-glucose conversion and no D-galactose conversion activity
H450G
the mutant shows a 3.6fold decrease in KM together with a 1.4fold increase in kcat for its substrate D-galactose and an overall improvement in the catalytic efficiency by a factor of 5 compared to the wild type enzyme
H450G
the mutant exhibits reduced D-glucose conversion and improved D-galactose conversion activity
H450G/V546C
mutant with increased activity with D-galactose, high activity with D-glucose, and considerably increased stability for the latter variant
H450G/V546C
the mutant exhibits reduced D-glucose conversion and improved D-galactose conversion activity
L545C
pre-steady-state kinetics of the oxidative half-reaction show higher kinetic constants as compared to the wild-type. The variant immobilized on glassy carbon electrodes displays high catalytic efficiency. L545C performs best among the variants studied
L547R
pre-steady-state kinetics of the oxidative half-reaction show higher kinetic constants as compared to the wild-type. The variant immobilized on glassy carbon electrodes displays high catalytic efficiency
Q448H
pre-steady-state kinetics of the oxidative half-reaction show higher kinetic constants as compared to the wild-type. The variant immobilized on glassy carbon electrodes displays high catalytic efficiency
T166R
pre-steady-state kinetics of the oxidative half-reaction show higher kinetic constants as compared to the wild-type. The variant immobilized on glassy carbon electrodes displays high catalytic efficiency
T169G
the FAD content of the mutant enzyme is similar to the wild type enzyme
T169G
the mutant exhibits reduced D-glucose conversion and D-galactose conversion activities
T169N
the FAD content of the mutant enzyme is similar to the wild type enzyme
T169S
the FAD content of the mutant enzyme is similar to the wild type enzyme
V546C
the mutant is characterized by elevated kcat values for both sugar substrates D-glucose and D-galactose, which is accompanied by elevated KM values
V546C
the mutant exhibits reduced D-glucose and D-galactose conversion activities
H167A
-
the mutant catalyzes the oxidation of D-glucose regiospecifically at the C2 position. The rate constant of flavin reduction decreases about 22fold compared to that of the wild type enzyme
H167A
-
site-directed mutagenesis, reductive activity of the mutant is highly reduced compared to the wild-type enzyme, the mutation ablates the covalent histidyl-FAD linkage. The H167A mutant enzyme oxidizes D-glucose regiospecifically at the C2 position, similarly to the wild-type enzyme, but noncovalent flavin in H167A mainly decreases the flavin reduction rate constant by 22fold
H167A
-
site-directed mutagenesis, reductive activity of the mutant is highly reduced compared to the wild-type enzyme, the mutation ablates the covalent histidyl-FAD linkage. The H167A mutant enzyme oxidizes D-glucose regiospecifically at the C2 position, similarly to the wild-type enzyme, but noncovalent flavin in H167A mainly decreases the flavin reduction rate constant by 22fold. In the crystal complex of the H167A mutant and 2FG (C3 oxidation), the substrate-recognition loop swings further away from the active site to assume the open conformation, whereas in the C2 oxidation complex of the H167A mutant and 3FG, the gating segment (residues 452-456) swings towards the active site to provide a binding pocket for the substrate
H167A
-
site-directed mutagenesis, the covalent linkag of the isoalloxazine ring of the cofactor FAD to His167 is destroyed, molecular dynamics, overview
H167A/H548R
-
site-directed mutagenesis, reductive activity of the mutant is reduced compared to the wild-type enzyme, contains noncovalently linked FAD, highest reductive activity at pH 10.5
H167A/H548R
-
site-directed mutagenesis, reductively inactive mutant, contains noncovalently linked FAD
T169A
-
the mutant shows severely decreased catalytic efficiency for D-glucose and D-galactose compared to the wild type enzyme
T169A
-
site-directed mutagenesis, the hydrogen bond between residue 169 and the N5 atom of FAD is absent, resulting in a change in the kinetic mechanism to one that consists of a ternary complex
T169A
-
site-directed mutagenesis, the hydrogen bond between Thr169 and the N5 atom of FAD is absent in the mutant.The kinetic mechanism of the T169A mutant with D-glucose or D-galactose indicates that a 2-keto-sugar product remains bound at the active site during the oxidative half-reaction
T169G
-
the mutant shows decreased catalytic efficiency for D-glucose and D-galactose compared to the wild type enzyme
T169N
-
the mutant shows decreased catalytic efficiency for D-glucose and increased catalytic efficiency for D-galactose compared to the wild type enzyme
T169S
-
the mutant shows increased kcat and Km values for D-glucose and D-galactose compared to the wild type enzyme
additional information
the deletion mutant DELTA454-456 shows 6fold lower kcat (D-galactose/O2), and 0.8fold elevated Km (D-galactose) value compared to the wild type enzyme
additional information
-
the deletion mutant DELTA454-456 shows 6fold lower kcat (D-galactose/O2), and 0.8fold elevated Km (D-galactose) value compared to the wild type enzyme
additional information
-
comparison the P2O active site in H167A, T169S or T169S-acetate complex with wild-type P2O-acetate complex, molecuar dynamics and spectroscopic analysi, detailed overview
additional information
-
mutagenesis of Phe454 and Tyr456 results in inactive enzymes, indicating that this region is functionally important for P2O
additional information
-
redox potential of wild-type and mutant enzymes, overview. Single mutants of H548 result in mixed populations of noncovalently bound and covalently linked FAD. Double mutants containing H167A are constructed, in which the covalent histidyl-FAD linkage is removed in addition to having the H548 mutation
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
stability of soluble and immobilized enzymes examined by incubating samples at 30°C for 7 days in citrate-phosphate buffer ranging from pH 4 to 8, similar stability obtained in free or immobilized form
687890
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4
under dry and wet storage conditions stable at 4°C, under dry storage conditions retainment of 100% of original activity after 2 days and of about 90% after one week, storage in working buffer solutions decreases activity by about 10% after one day and by more than 50% after one week
60
T169G/E542K/V546C mutant, half life is increased 76fold compared to wild-type
60 - 70
69
melting temperature of mutant L547W, increased half-life compared to wild-type
70
T169G/E542K/V546C mutant, half life is increased 350fold compared to wild-type
71.1
melting temperature of mutant L547G, increased half-life compared to wild-type
74.2
melting temperature of mutant E542K, increased half-life compared to wild-type
74.3
melting temperature of mutant E542R, increased half-life compared to wild-type
additional information
60 - 70
the half life of the wild type enzyme at 60°C and 70°C is 12 min and 0.07 min, respectively. The melting temperature of the wild type enzyme is at 63.5°C
60 - 70
the wild type enzyme shows a half life of 10 min at 60°C and of less than 1 min at 70°C
additional information
mutant L537G/E542K, L537G/E542R, L537W/E542K and L537W/E542R show more complex melting curves, increased half-life compared to wild-type
additional information
-
mutant L537G/E542K, L537G/E542R, L537W/E542K and L537W/E542R show more complex melting curves, increased half-life compared to wild-type
additional information
-
thermal inactivation of free and immobilized enzyme activity investigated at 40, 50, 60 and 70°C in 50 mM phosphate buffer at pH 6.0 over 7 days, stability of immobilized enzyme shown to be significantly higher at 60°C, no stabilizing effect observed at 70°C
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni2+ immobilized column with Chelating Sepharose Fast Flow combined with Äkta Purifier System, concentrated in an Amicon Ultra Centrifugal Filter Device
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinit chromatography and ulrafiltration
protein purification by immobilized metal affinity chromatography, ultrafiltration, concentration of enzymes
-
recmbinant enzyme from Escherichia coli by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
-
stored, wet cell paste is treated with lysozyme, PMSF, EDTA buffer. Following steps are RNAse and DNAse treatment, sonication, dialyse, DEAE Sepharose Fast Flow column, dialyse, Phenyl Sepharose Fast Flow column, concentration by ammonium sulfate saturation, final dialyse. Stored at 20°C
-
using ammonium sulfate fractionation, hydrophobic interaction, ion-exchange chromatography, and gel filtration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli BL21-DE3, recombinant host strains shown to be different in formation of inclusion bodies between inserts of Trametes ochracea and Trametes pubescens at higher growth temperatures, plasmids pSE33 and pRQ5 used for site-directed recombination
expressed in Escherichia coli BL21-DE3, recombinant protein, plasmids pHL2e and pCL22 encoding the E542K mutant used
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
expressed in Escherichia coli BL21-DE3, recombinant protein, expression vector pET21
-
recombinant expression of pyranose 2-oxidase in Escherichia coli strain Rosetta(DE3)pLysS
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
biotechnology
energy production
-
enzyme P2O has the potential to be useful for biofuel cell applications
synthesis
-
enzyme P2O is a useful biocatalyst in several biotechnological applications, including biotransformation of carbohydrates such as D-glucose and D-galactose to generate 2-oxo-sugars that can be further reduced at the C1 position to yield D-fructose and D-tagatose, respectively
biotechnology
biocatalyst for carbohydrate transformations toward higher-value products
biotechnology
-
studies on stabilization of enzymatic activity by immobilization
biotechnology
-
enzyme P2O is a useful biocatalyst in several biotechnological applications, including biotransformation of carbohydrates such as D-glucose and D-galactose to generate 2-oxo-sugars that can be further reduced at the C1 position to yield D-fructose and D-tagatose, respectively
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Leitner, C.; Volc, J.; Haltrich, D.
Purification and characterization of pyranose oxidase from the white rot fungus Trametes multicolor
Appl. Environ. Microbiol.
67
3636-3644
2001
Trametes ochracea
Halada, P.; Leitner, C.; Sedmera, P.; Haltrich, D.; Volc, J.
Identification of the covalent flavin adenine dinucleotide-binding region in pyranose 2-oxidase from Trametes multicolor
Anal. Biochem.
314
235-242
2003
Trametes ochracea
Hallberg, B.M.; Leitner, C.; Haltrich, D.; Divne, C.
Crystal structure of the 270 kDa homotetrameric lignin-degrading enzyme pyranose 2-oxidase
J. Mol. Biol.
341
781-796
2004
Trametes ochracea
Kujawa, M.; Ebner, H.; Leitner, C.; Hallberg, B.M.; Prongjit, M.; Sucharitakul, J.; Ludwig, R.; Rudsander, U.; Peterbauer, C.; Chaiyen, P.; Haltrich, D.; Divne, C.
Structural basis for substrate binding and regioselective oxidation of monosaccharides at C3 by pyranose 2-oxidase
J. Biol. Chem.
281
35104-35115
2006
Trametes ochracea (Q7ZA32)
Tasca, F.; Timur, S.; Ludwig, R.; Haltrich, D.; Volc, J.; Antiochia, R.; Gorton, L.
Amperometric biosensors for detection of sugars based on the electrical wiring of different pyranose oxidases and pyranose dehydrogenases with osmium redox polymer on graphite electrodes
Electroanalysis
19
294-302
2007
Trametes ochracea (Q7ZA32)
-
Maresova, H.; Palyzova, A.; Kyslik, P.
The C-terminal region controls correct folding of genus Trametes pyranose 2-oxidases
J. Biotechnol.
130
229-235
2007
Trametes pubescens (Q5G234), Trametes pubescens, Trametes ochracea (Q7ZA32), Trametes ochracea
Sukyai, P.; Rezic, T.; Lorenz, C.; Mueangtoom, K.; Lorenz, W.; Haltrich, D.; Ludwig, R.
Comparing soluble and co-immobilized catalysts for 2-ketoaldose production by pyranose 2-oxidase and auxiliary enzymes
J. Biotechnol.
135
281-290
2008
Trametes ochracea
Rungsrisuriyachai, K.; Gadda, G.
A pH switch affects the steady-state kinetic mechanism of pyranose 2-oxidase from Trametes ochracea
Arch. Biochem. Biophys.
483
10-15
2009
Trametes ochracea
Prongjit, M.; Sucharitakul, J.; Wongnate, T.; Haltrich, D.; Chaiyen, P.
Kinetic mechanism of pyranose 2-oxidase from trametes multicolor
Biochemistry
48
4170-4180
2009
Trametes ochracea
Van Hecke, W.; Salaheddin, C.; Ludwig, R.; Dewulf, J.; Haltrich, D.; Van Langenhove, H.
Biocatalytic cascade oxidation using laccase for pyranose 2-oxidase regeneration
Biores. Technol.
100
5566-5573
2009
Trametes ochracea, Trametes ochracea MB 49
Spadiut, O.; Radakovits, K.; Pisanelli, I.; Salaheddin, C.; Yamabhai, M.; Tan, T.C.; Divne, C.; Haltrich, D.
A thermostable triple mutant of pyranose 2-oxidase from Trametes multicolor with improved properties for biotechnological applications
Biotechnol. J.
4
525-534
2009
Trametes ochracea (Q7ZA32), Trametes ochracea
Salaheddin, C.; Spadiut, O.; Ludwig, R.; Tan, T.C.; Divne, C.; Haltrich, D.; Peterbauer, C.
Probing active-site residues of pyranose 2-oxidase from Trametes multicolor by semi-rational protein design
Biotechnol. J.
4
535-543
2009
Trametes ochracea (Q7ZA32), Trametes ochracea
Spadiut, O.; Leitner, C.; Salaheddin, C.; Varga, B.; Vertessy, B.G.; Tan, T.C.; Divne, C.; Haltrich, D.
Improving thermostability and catalytic activity of pyranose 2-oxidase from Trametes multicolor by rational and semi-rational design
FEBS J.
276
776-792
2009
Trametes ochracea (Q7ZA32), Trametes ochracea
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
J. Biotechnol.
139
250-257
2009
Trametes ochracea
Sucharitakul, J.; Wongnate, T.; Chaiyen, P.
Kinetic isotope effects on the noncovalent flavin mutant protein of pyranose 2-oxidase reveal insights into the flavin reduction mechanism
Biochemistry
49
3753-3765
2010
Trametes ochracea
Spadiut, O.; Tan, T.C.; Pisanelli, I.; Haltrich, D.; Divne, C.
Importance of the gating segment in the substrate-recognition loop of pyranose 2-oxidase
FEBS J.
277
2892-2909
2010
Trametes ochracea (Q7ZA32), Trametes ochracea
Spadiut, O.; Nguyen, T.T.; Haltrich, D.
Thermostable variants of pyranose 2-oxidase showing altered substrate selectivity for glucose and galactose
J. Agric. Food Chem.
58
3465-3471
2010
Trametes ochracea (Q7ZA32), Trametes ochracea
Pitsawong, W.; Sucharitakul, J.; Prongjit, M.; Tan, T.C.; Spadiut, O.; Haltrich, D.; Divne, C.; Chaiyen, P.
A conserved active-site threonine is important for both sugar and flavin oxidations of pyranose 2-oxidase
J. Biol. Chem.
285
9697-9705
2010
Trametes ochracea
Tan, T.C.; Pitsawong, W.; Wongnate, T.; Spadiut, O.; Haltrich, D.; Chaiyen, P.; Divne, C.
H-bonding and positive charge at the N5/O4 locus are critical for covalent flavin attachment in trametes pyranose 2-oxidase
J. Mol. Biol.
402
578-594
2010
Trametes ochracea (Q7ZA32), Trametes ochracea
Prongjit, M.; Sucharitakul, J.; Palfey, B.A.; Chaiyen, P.
Oxidation mode of pyranose 2-oxidase is controlled by pH
Biochemistry
52
1437-1445
2013
Trametes ochracea
Wongnate, T.; Sucharitakul, J.; Chaiyen, P.
Identification of a catalytic base for sugar oxidation in the pyranose 2-oxidase reaction
ChemBioChem
12
2577-2586
2011
Trametes ochracea
Wongnate, T.; Chaiyen, P.
The substrate oxidation mechanism of pyranose 2-oxidase and other related enzymes in the glucose-methanol-choline superfamily
FEBS J.
280
3009-3027
2013
Trametes ochracea
Decamps, K.; Joye, I.; Haltrich, D.; Nicolas, J.; Courtin, C.; Delcour, J.
Biochemical characteristics of Trametes multicolor pyranose oxidase and Aspergillus niger glucose oxidase and implications for their functionality in wheat flour dough
Food Chem.
131
1485-1492
2012
Trametes ochracea (Q7ZA32), Trametes ochracea MB49 (Q7ZA32)
Tan, T.C.; Haltrich, D.; Divne, C.
Regioselective control of beta-D-glucose oxidation by pyranose 2-oxidase is intimately coupled to conformational degeneracy
J. Mol. Biol.
409
588-600
2011
Trametes ochracea (Q7ZA32), Trametes ochracea
Taniguchi, S.; Chosrowjan, H.; Wongnate, T.; Sucharitakul, J.; Chaiyen, P.; Tanaka, F.
Ultrafast fluorescence dynamics of flavin adenine dinucleotide in pyranose 2-oxidases variants and their complexes with acetate: conformational heterogeneity with different dielectric constants
J. Photochem. Photobiol. A
245
33-42
2012
Trametes ochracea
-
Decamps, K.; Gryp, G.; Joye, I.J.; Courtin, C.M.; Delcour, J.A.
Impact of pyranose oxidase from Trametes multicolor, glucose oxidase from Aspergillus niger and hydrogen peroxide on protein agglomeration in wheat flour gluten-starch separation
Food Chem.
148
235-239
2014
Trametes ochracea (Q7ZA32), Trametes ochracea
Decamps, K.; Joye, I.J.; Rakotozafy, L.; Nicolas, J.; Courtin, C.M.; Delcour, J.A.
The bread dough stability improving effect of pyranose oxidase from Trametes multicolor and glucose oxidase from Aspergillus niger: unraveling the molecular mechanism
J. Agric. Food Chem.
61
7848-7854
2013
Trametes ochracea (Q7ZA32), Trametes ochracea
Lugsanangarm, K.; Nueangaudom, A.; Kokpol, S.; Pianwanit, S.; Nunthaboot, N.; Tanaka, F.; Taniguchi, S.; Chosrowjan, H.
Heterogeneous subunit structures in the pyranose 2-oxidase homotetramer revealed by theoretical analysis of the rates of photoinduced electron transfer from a tryptophan to the excited flavin
J. Photochem. Photobiol. A
306
469-476
2015
Trametes ochracea (Q7ZA32)
-
Brugger, D.; Suetzl, L.; Zahma, K.; Haltrich, D.; Peterbauer, C.K.; Stoica, L.
Electrochemical characterization of the pyranose 2-oxidase variant N593C shows a complete loss of the oxidase function with full preservation of substrate (dehydrogenase) activity
Phys. Chem. Chem. Phys.
18
32072-32077
2016
Trametes ochracea
Halada, P.; Brugger, D.; Volc, J.; Peterbauer, C.K.; Leitner, C.; Haltrich, D.
Oxidation of Phe454 in the gating segment inactivates Trametes multicolor pyranose oxidase during substrate turnover
PLoS ONE
11
e0148108
2016
Trametes ochracea
Brugger, D.; Krondorfer, I.; Shelswell, C.; Huber-Dittes, B.; Haltrich, D.; Peterbauer, C.K.
Engineering pyranose 2-oxidase for modified oxygen reactivity
PLoS ONE
9
e109242
2014
Trametes ochracea (Q7ZA32)
Tan, T.C.; Spadiut, O.; Gandini, R.; Haltrich, D.; Divne, C.
Structural basis for binding of fluorinated glucose and galactose to Trametes multicolor pyranose 2-oxidase variants with improved galactose conversion
PLoS ONE
9
e86736
2014
Trametes ochracea (Q7ZA32), Trametes ochracea
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