Literature summary for 1.1.3.10 extracted from

Brugger, D.; Krondorfer, I.; Shelswell, C.; Huber-Dittes, B.; Haltrich, D.; Peterbauer, C.K.
Engineering pyranose 2-oxidase for modified oxygen reactivity (2014), PLoS ONE, 9, e109242.

Cloned(Commentary)

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

Protein Variants

Protein Variants	Comment	Organism
L545C	the mutant shows 19% of wild type activity	Trametes ochracea
L547R	the mutant shows 0.99% of wild type activity	Trametes ochracea
N593C	the mutant shows 0.058% of wild type activity	Trametes ochracea
Q448H	the mutant shows 1.5% of wild type activity	Trametes ochracea
T166R	the mutant shows 39% of wild type activity	Trametes ochracea

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.018	-	D-glucose	mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.042	-	D-glucose	mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.046	-	D-glucose	mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.057	-	D-glucose	mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.082	-	D-glucose	mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.092	-	D-glucose	mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.12	-	D-glucose	mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.13	-	D-glucose	wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.14	-	D-glucose	mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.14	-	D-glucose	mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.18	-	D-glucose	mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.23	-	D-glucose	mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.24	-	D-glucose	mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.24	-	D-glucose	wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.25	-	D-glucose	wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.4	-	D-glucose	mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.56	-	D-glucose	mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.72	-	D-glucose	mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.81	-	D-glucose	mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.98	-	D-glucose	mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1.6	-	D-glucose	mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1.7	-	D-glucose	mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1.9	-	D-glucose	wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
2.1	-	D-glucose	mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
65000	-	x * 65000, SDS-PAGE	Trametes ochracea

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-glucose + O2	Trametes ochracea	-	2-dehydro-D-glucose + H2O2	-	?

Organism

Organism	UniProt	Comment	Textmining
Trametes ochracea	Q7ZA32	-	-

Purification (Commentary)

Purification (Comment)	Organism
IMAC Ni-charged resin column chromatography	Trametes ochracea

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
D-glucose + 1,4-benzoquinone	-	Trametes ochracea	2-dehydro-D-glucose + 1,4-hydroquinone	-	?
D-glucose + 2,6-dichlorophenolindophenol	-	Trametes ochracea	2-dehydro-D-glucose + reduced 2,6-dichlorophenolindophenol	-	?
D-glucose + ferricenium ion	-	Trametes ochracea	2-dehydro-D-glucose + ferrocenium ion	-	?
D-glucose + O2	-	Trametes ochracea	2-dehydro-D-glucose + H2O2	-	?

Subunits

Subunits	Comment	Organism
?	x * 65000, SDS-PAGE	Trametes ochracea

Synonyms

Synonyms	Comment	Organism
glucose 2-oxidase	-	Trametes ochracea
POX	-	Trametes ochracea
pyranose 2-Oxidase	-	Trametes ochracea
pyranose:oxygen 2-oxidoreductase	-	Trametes ochracea

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.18	-	D-glucose	mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
0.38	-	D-glucose	mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1.1	-	D-glucose	mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
2.7	-	D-glucose	mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
3.9	-	D-glucose	mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
4.6	-	D-glucose	mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
8.1	-	D-glucose	mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
11	-	D-glucose	mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
12	-	D-glucose	mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
22	-	D-glucose	mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
26	-	D-glucose	mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
27	-	D-glucose	mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
42	-	D-glucose	wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
50	-	D-glucose	mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
70	-	D-glucose	wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
110	-	D-glucose	mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
134	-	D-glucose	mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
134	-	D-glucose	mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
142	-	D-glucose	mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
151	-	D-glucose	wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
152	-	D-glucose	wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
166	-	D-glucose	mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
265	-	D-glucose	mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
442	-	D-glucose	mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Trametes ochracea

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.18	-	D-glucose	mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
2.8	-	D-glucose	mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
4.6	-	D-glucose	mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
22	-	D-glucose	mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
27	-	D-glucose	mutant enzyme N593C, with 1 ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
32	-	D-glucose	mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
34	-	D-glucose	mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
38	-	D-glucose	wild type enzyme, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
48	-	D-glucose	mutant enzyme Q448H, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
58	-	D-glucose	mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
70	-	D-glucose	mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
79	-	D-glucose	mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
120	-	D-glucose	mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
127	-	D-glucose	mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
203	-	D-glucose	mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
281	-	D-glucose	mutant enzyme T166R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
308	-	D-glucose	wild type enzyme, with O2 as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
584	-	D-glucose	mutant enzyme L547R, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
592	-	D-glucose	wild type enzyme, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
632	-	D-glucose	wild type enzyme, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1193	-	D-glucose	mutant enzyme L545C, with ferrocenium ion as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1213	-	D-glucose	mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
1465	-	D-glucose	mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea
2608	-	D-glucose	mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C	Trametes ochracea