BRENDA - Enzyme Database

Heterologous overexpression of Glomerella cingulata FAD-dependent glucose dehydrogenase in Escherichia coli and Pichia pastoris

Sygmund, C.; Staudigl, P.; Klausberger, M.; Pinotsis, N.; Djinovic-Carugo, K.; Gorton, L.; Haltrich, D.; Ludwig, R.; Microb. Cell Fact. 10, 106 (2011)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.5.9
recombinant overexpression in Escherichia coli strains Rosetta 2, T7 Express and T7 Express (pGro7) and Pichia pastoris strain X-33, with a much higher expression level and 4800fold higher enzyme activity in Pichia pastoris, fed-batch cultivation of a Pichia pastoris transformant, method evaluation, overview
Colletotrichum gloeosporioides
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.5.9
2
3
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
10.1
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
10.2
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.1
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
19
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
21
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
24
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
26
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.5.9
67000
-
x * 67000, deglycosylated recombinant enzyme expressed from Pichia pastoris, SDS-PAGE
Colletotrichum gloeosporioides
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.5.9
D-glucose + a quinone
Colletotrichum gloeosporioides
-
D-glucono-1,5-lactone + a quinol
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.5.9
Colletotrichum gloeosporioides
-
anamorph Colletotrichum gloeosporoides
-
Posttranslational Modification
EC Number
Posttranslational Modification
Commentary
Organism
1.1.5.9
glycoprotein
-
Colletotrichum gloeosporioides
Purification (Commentary)
EC Number
Commentary
Organism
1.1.5.9
recombinant enzyme from Pichia pastoris strain X-33 5.1fold by hydrophobic interaction and anion exchange chromatography
Colletotrichum gloeosporioides
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.1.5.9
836
-
purified recombinant enzyme expressed from Pichia pastoris, pH 7.5, 30C
Colletotrichum gloeosporioides
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.5.9
D-glucose + a quinone
-
725879
Colletotrichum gloeosporioides
D-glucono-1,5-lactone + a quinol
-
-
-
?
1.1.5.9
D-glucose + ferricenium ion
-
725879
Colletotrichum gloeosporioides
D-glucono-1,5-lactone + ferrocenium ion
-
-
-
?
1.1.5.9
D-xylose + ferricenium ion
-
725879
Colletotrichum gloeosporioides
D-xylono-1,5-lactone + ferrocenium ion
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.5.9
?
x * 67000, deglycosylated recombinant enzyme expressed from Pichia pastoris, SDS-PAGE
Colletotrichum gloeosporioides
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.5.9
46
-
recombinant enzyme expressed from Pichia pastoris
Colletotrichum gloeosporioides
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.1.5.9
56
-
pH-dependent thermal stability with the highest Tm values in the acidic range of pH 4.5 to pH 6.4. The maximum Tm value of 56C is measured in 50 mM sodium acetate buffer pH 5.0 and in 50 mM MES buffer pH 5.8
Colletotrichum gloeosporioides
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.5.9
40
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
53
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
60
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
61
-
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
179
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
180
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
380
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
418
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.5.9
7.5
-
recombinant enzyme
Colletotrichum gloeosporioides
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
1.1.5.9
5
5.8
pH-dependent thermal stability with the highest Tm values in the acidic range of pH 4.5 to pH 6.4. The maximum Tm value of 56C is measured in 50 mM sodium acetate buffer pH 5.0 and in 50 mM MES buffer pH 5.8
Colletotrichum gloeosporioides
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.5.9
FAD
dependent on
Colletotrichum gloeosporioides
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.5.9
recombinant overexpression in Escherichia coli strains Rosetta 2, T7 Express and T7 Express (pGro7) and Pichia pastoris strain X-33, with a much higher expression level and 4800fold higher enzyme activity in Pichia pastoris, fed-batch cultivation of a Pichia pastoris transformant, method evaluation, overview
Colletotrichum gloeosporioides
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.5.9
FAD
dependent on
Colletotrichum gloeosporioides
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.5.9
2
3
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
10.1
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
10.2
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.1
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
19
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
21
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
24
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
26
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.5.9
67000
-
x * 67000, deglycosylated recombinant enzyme expressed from Pichia pastoris, SDS-PAGE
Colletotrichum gloeosporioides
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.5.9
D-glucose + a quinone
Colletotrichum gloeosporioides
-
D-glucono-1,5-lactone + a quinol
-
-
?
Posttranslational Modification (protein specific)
EC Number
Posttranslational Modification
Commentary
Organism
1.1.5.9
glycoprotein
-
Colletotrichum gloeosporioides
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.5.9
recombinant enzyme from Pichia pastoris strain X-33 5.1fold by hydrophobic interaction and anion exchange chromatography
Colletotrichum gloeosporioides
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.1.5.9
836
-
purified recombinant enzyme expressed from Pichia pastoris, pH 7.5, 30C
Colletotrichum gloeosporioides
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.5.9
D-glucose + a quinone
-
725879
Colletotrichum gloeosporioides
D-glucono-1,5-lactone + a quinol
-
-
-
?
1.1.5.9
D-glucose + ferricenium ion
-
725879
Colletotrichum gloeosporioides
D-glucono-1,5-lactone + ferrocenium ion
-
-
-
?
1.1.5.9
D-xylose + ferricenium ion
-
725879
Colletotrichum gloeosporioides
D-xylono-1,5-lactone + ferrocenium ion
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.5.9
?
x * 67000, deglycosylated recombinant enzyme expressed from Pichia pastoris, SDS-PAGE
Colletotrichum gloeosporioides
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.5.9
46
-
recombinant enzyme expressed from Pichia pastoris
Colletotrichum gloeosporioides
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.1.5.9
56
-
pH-dependent thermal stability with the highest Tm values in the acidic range of pH 4.5 to pH 6.4. The maximum Tm value of 56C is measured in 50 mM sodium acetate buffer pH 5.0 and in 50 mM MES buffer pH 5.8
Colletotrichum gloeosporioides
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.5.9
40
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
53
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
60
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
61
-
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
179
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
180
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
380
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
418
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.5.9
7.5
-
recombinant enzyme
Colletotrichum gloeosporioides
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
1.1.5.9
5
5.8
pH-dependent thermal stability with the highest Tm values in the acidic range of pH 4.5 to pH 6.4. The maximum Tm value of 56C is measured in 50 mM sodium acetate buffer pH 5.0 and in 50 mM MES buffer pH 5.8
Colletotrichum gloeosporioides
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.5.9
1.9
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2.5
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2.7
-
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.6
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.7
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
20
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
24.5
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.5.9
1.9
-
D-xylose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2
-
D-xylose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2.5
-
D-xylose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
2.7
-
D-xylose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.6
-
D-glucose
native enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
17.7
-
D-glucose
recombinant enzyme, pH 5.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
20
-
D-glucose
native enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides
1.1.5.9
24.5
-
D-glucose
recombinant enzyme, pH 7.5, 30C
Colletotrichum gloeosporioides