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Literature summary for 1.1.3.10 extracted from

  • Spadiut, O.; Nguyen, T.T.; Haltrich, D.
    Thermostable variants of pyranose 2-oxidase showing altered substrate selectivity for glucose and galactose (2010), J. Agric. Food Chem., 58, 3465-3471.
    View publication on PubMed

Application

Application Comment Organism
biotechnology biocatalyst for carbohydrate transformations toward higher-value products Trametes ochracea

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
E542K the mutant is characterized by reduced KM values for both D-glucose and D-galactose and significantly increased stability Trametes ochracea
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 Trametes ochracea
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 Trametes ochracea
H450G/V546C mutant with increased activity with D-galactose, high activity with D-glucose, and considerably increased stability for the latter variant Trametes ochracea
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 Trametes ochracea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.521
-
D-glucose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.77
-
D-glucose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.987
-
D-glucose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.99
-
D-glucose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
2.43
-
D-glucose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
2.45
-
D-galactose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
3.06
-
D-glucose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
3.87
-
D-galactose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
7.8
-
D-galactose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
8.79
-
D-galactose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
12
-
D-galactose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
46.2
-
D-galactose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea

Organism

Organism UniProt Comment Textmining
Trametes ochracea Q7ZA32
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Trametes ochracea

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose + O2 His-tagged recombinant wild type enzyme strongly prefers D-glucose to D-galactose as its substrate Trametes ochracea 2-dehydro-D-galactose + H2O2
-
?
D-glucose + O2 His-tagged recombinant wild type enzyme strongly prefers D-glucose to D-galactose as its substrate Trametes ochracea 2-dehydro-D-glucose + H2O2
-
?

Subunits

Subunits Comment Organism
homotetramer
-
Trametes ochracea

Synonyms

Synonyms Comment Organism
glucose 2-oxidase
-
Trametes ochracea
P2Ox
-
Trametes ochracea
pyranose 2-Oxidase
-
Trametes ochracea
pyranose:oxygen 2-oxidoreductase
-
Trametes ochracea

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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 Trametes ochracea

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.51
-
D-galactose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
2.59
-
D-galactose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
2.84
-
D-galactose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
3.51
-
D-galactose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
5.92
-
D-galactose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
6.57
-
D-galactose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
6.81
-
D-glucose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
12.5
-
D-glucose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
16.8
-
D-glucose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
35.9
-
D-glucose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
48.1
-
D-glucose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
88.6
-
D-glucose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°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 Structure
0.142
-
D-galactose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.286
-
D-galactose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.364
-
D-galactose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.493
-
D-galactose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
0.669
-
D-galactose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
1.43
-
D-galactose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
8.83
-
D-glucose mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
12.7
-
D-glucose mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
13.5
-
D-glucose mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
29
-
D-glucose mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
51.2
-
D-glucose His-tagged recombinant wild type enzyme, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea
68.9
-
D-glucose mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C Trametes ochracea