Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.3.10 extracted from

  • Bastian, S.; Rekowski, M.J.; Witte, K.; Heckmann-Pohl, D.M.; Giffhorn, F.
    Engineering of pyranose 2-oxidase from Peniophora gigantea towards improved thermostability and catalytic efficiency (2005), Appl. Microbiol. Biotechnol., 67, 654-663.
    View publication on PubMed
Search for more literature for 1.1.3.10

Application

Application Comment Organism
synthesis application of the engineered enzyme K312E in bioconversion of L-sorbose to 5-keto-D-fructose Phlebiopsis gigantea

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3), yielding an intracellular and enzymatically active P2OxB with a volumetric yield of 500 units/ml Phlebiopsis gigantea

Protein Variants

Protein Variants Comment Organism
E540K mutant with increased thermo- and pH-stability compared with wild-type, concomitantly with increased catalytic efficiencies (turnover number/KM-value) for D-xylose and L-sorbose Phlebiopsis gigantea
K312E mutant enzyme with significant improvements with respect to the ratio of turnover number to Km-value for D-glucose (5.3fold), methyl-beta-D-glucoside (2.0fold), D-galactose (4.8fold), D-xylose (59.9fold), and L-sorbose (69.9fold) compared with wild-type P2Ox Phlebiopsis gigantea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.4
-
 D-glucose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
0.6
-
 D-glucose pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
0.6
-
 D-glucose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
0.8
-
 D-glucose pH 7.3, 22°C, recombinant wild-type enzyme Phlebiopsis gigantea
1.1
-
 D-glucose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
2.4
-
 D-xylose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
2.9
-
 L-sorbose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
5
-
 L-sorbose pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
5.2
-
 D-galactose pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
5.4
-
 L-sorbose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
6.1
-
 D-galactose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
6.3
-
 D-galactose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
6.6
-
 D-xylose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
7.9
-
 D-xylose pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
8.2
-
 D-galactose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
11.3
-
 2-deoxy-D-glucose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
22.1
-
 D-allose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
22.3
-
 D-allose pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
25
-
 D-xylose pH 7.3, 22°C, recombinant wild-type enzyme Phlebiopsis gigantea
29.4
-
 D-xylose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
47
-
 L-sorbose pH 7.3, 22°C, recombinant wild-type enzyme Phlebiopsis gigantea
50
-
 methyl-beta-D-glucoside pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
50
-
 L-sorbose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
55.7
-
 D-allose pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea
68
-
 D-allose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
79
-
 2-deoxy-D-glucose pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
99.3
-
 2-deoxy-D-glucose pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
256
-
 methyl-beta-D-glucoside pH 7.3, 22°C, mutant enzyme E540K Phlebiopsis gigantea
261
-
 methyl-beta-D-glucoside pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea
289
-
 methyl-beta-D-glucoside pH 7.3, 22°C, wild-type enzyme Phlebiopsis gigantea

Organism

Organism UniProt Comment Textmining
Phlebiopsis gigantea Q6UG02
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes Phlebiopsis gigantea

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.74
-
 recombinant wild-type enzyme Phlebiopsis gigantea
13.4
-
 mutant enzyme E540K with a C-terminal His6-tag Phlebiopsis gigantea

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-deoxy-D-glucose + O2
-
 Phlebiopsis gigantea 2-deoxy-3-dehydro-D-glucose + H2O2
-
 r
D-allose + O2
-
 Phlebiopsis gigantea ?
-
 ?
D-galactose + O2
-
 Phlebiopsis gigantea ?
-
 ?
D-glucose + O2
-
 Phlebiopsis gigantea 2-dehydro-D-glucose + H2O2
-
 ?
D-xylose + O2
-
 Phlebiopsis gigantea ?
-
 ?
L-sorbose + O2
-
 Phlebiopsis gigantea ?
-
 ?
methyl-beta-D-glucoside + O2
-
 Phlebiopsis gigantea ?
-
 ?

Synonyms

Synonyms Comment Organism
P2Ox
-
 Phlebiopsis gigantea

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 wild-type enzyme Phlebiopsis gigantea
50
-
 mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 58 30°C: about 60% of maximal, 58°C: about 70% of maximal activity, mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea
30 53 30°C: about 60% of maximal activity, 58°C: about 75% of maximal activity Phlebiopsis gigantea

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
 pH 7.0, 30 min, wild-type enzyme, mutant enzyme K312E with a C-terminal His6-tag and mutant enzyme E540K are stable Phlebiopsis gigantea
50
-
 pH 7.0, 30 min, wild-type enzyme loses about 40% of its activity, mutant enzyme K312E with a C-terminal His6-tag and mutant enzyme E540K are stable Phlebiopsis gigantea
55
-
 pH 7.0, 30 min, wild-type enzyme loses 95% of its activity, mutant enzyme K312E with a C-treminal His6-tag and mutant enzyme E540K lose about 10% of its activity Phlebiopsis gigantea
60
-
 pH 7.0, 30 min, wild-type enzyme completely loses its activity, mutant enzyme K312E with a C-terminal His6-tag loses about 40% of its activity and mutant enzyme E540K loses about 10% of its activity Phlebiopsis gigantea

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5 6 mutant enzyme E540K Phlebiopsis gigantea
6
-
 mutant enzyme K312E with a C-terminal His6-tag Phlebiopsis gigantea

pH Stability

pH Stability pH Stability Maximum Comment Organism
1 2 pH 7.0, 30 min, wild-type enzyme loses about 75% of its activity, mutant enzyme K312E with a C-terminal His6-tag loses about 70% of its activity and mutant enzyme E540K loses about 10% of its activity Phlebiopsis gigantea
6
-
 30 min, 30 min, wild-type enzyme and mutant enzyme E540K are stable, mutant enzyme K312E with a C-terminal His6-tag loses about 10% of its activity Phlebiopsis gigantea
7 9 30 min, wild-type enzyme, mutant enzyme K312E with a C-terminal His6-tag and mutant enzyme E540K are stable Phlebiopsis gigantea
10
-
 pH 7.0, 30 min, wild-type enzyme loses about 20% of its activity, mutant enzyme K312E with a C-terminal His6-tag and mutant enzyme E540K are stable Phlebiopsis gigantea
11
-
 pH 7.0, 30 min, wild-type enzyme loses about 40% of its activity, mutant enzyme K312E with a C-terminal His6-tag loses about 10% of its activity, mutant enzyme E540K is stable Phlebiopsis gigantea