BRENDA - Enzyme Database

Characterization of 2,4-dinitrotoluene dioxygenase from recombinant Escherichia coli strain PFJS39: Its direct interaction with vitreoscilla hemoglobin

Khleifat, K.M.; Bioremediat. J. 14, 38-53 (2010)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.14.12.24
2-mercaptoethanol
required for maximal activity, 292.3% activity at 1% (v/v)
Burkholderia sp.
1.14.12.24
ATP
124% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
FMN
230.7% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
GTP
116% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
lipase
176% activity at 1 mg per g wet weight of cells
Burkholderia sp.
1.14.12.24
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
1.14.12.24
phosphatidylcholine
125% activity at 0.05 mg
Burkholderia sp.
1.14.12.24
phosphatidylethanolamine
144% activity at 0.1 mg per g wet weight of cells
Burkholderia sp.
1.14.12.24
Phospholipase A
125% activity at 0.01 mg
Burkholderia sp.
Cloned(Commentary)
EC Number
Commentary
Organism
1.14.12.24
expressed in Escherichia coli PFJS39 cells
Burkholderia sp.
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.12.24
deoxycholate
about 68% residual activity at 6% (v/v)
Burkholderia sp.
1.14.12.24
Dodecyl sulfate
about 40% residual activity at 1% (v/v)
Burkholderia sp.
1.14.12.24
FAD
-
Burkholderia sp.
1.14.12.24
spermidine
80% residual activity at 0.05 mg
Burkholderia sp.
1.14.12.24
Triton X-100
about 50% residual activity at 2% (v/v)
Burkholderia sp.
1.14.12.24
Tween 20
about 36% residual activity at 2% (v/v)
Burkholderia sp.
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.12.24
0.05
-
O2
at pH 7.0 and 37°C
Burkholderia sp.
1.14.12.24
0.18
-
2,4-dinitrotoluene
at pH 7.0 and 37°C
Burkholderia sp.
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.12.24
Fe2+
required for activity, 238.5% activity at 1 mM
Burkholderia sp.
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.12.24
100000
-
gel filtration
Burkholderia sp.
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp.
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp. DNT
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.12.24
Burkholderia sp.
-
-
-
1.14.12.24
Burkholderia sp. DNT
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.12.24
ammonium sulfate precipitation, column chromatography, and Sephadex G-100 gel filtration
Burkholderia sp.
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.14.12.24
2.4
-
crude extract, at pH 7.0 and 37°C
Burkholderia sp.
1.14.12.24
37.3
-
after 15.7fold, at pH 7.0 and 37°C
Burkholderia sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp.
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp. DNT
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.12.24
heterotrimer
1 * 52000-71000, 1 * 23000-25500, 1 * 12000-16500, SDS-PAGE
Burkholderia sp.
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.12.24
37
-
-
Burkholderia sp.
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
1.14.12.24
4
45
activity rapidly diminishes when the enzyme is heated above 45°C. At 20°C, the activity is decreased about 25% compared with 37°C. At 15°C, the activity decreases about 85% compared with 37°C, and at 4-5°C, the activity is totally absent
Burkholderia sp.
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.14.12.24
-
4
the enzyme shows 5% to 15% loss in catalytic activity when left at 0-4°C for several weeks
Burkholderia sp.
1.14.12.24
25
37
the enzyme is stable for 2 days when incubated with substrate at 37°C, 30°C, and 25°C
Burkholderia sp.
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.12.24
7
-
-
Burkholderia sp.
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.14.12.24
4
10
the enzyme is stable when incubated at pH 6 and 7, but its optimal activity is at pH 7.0. The activity is decreased significantly when incubated at pH 4.0, 5.0, 8.0, and 10.0
Burkholderia sp.
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.12.24
FAD
-
Burkholderia sp.
1.14.12.24
NAD+
2fold enhancement of activity in the presence of NAD+
Burkholderia sp.
1.14.12.24
NADH
5fold enhancement of activity in the presence of NADH
Burkholderia sp.
1.14.12.24
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.14.12.24
2-mercaptoethanol
required for maximal activity, 292.3% activity at 1% (v/v)
Burkholderia sp.
1.14.12.24
ATP
124% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
FMN
230.7% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
GTP
116% activity at 0.1 mM
Burkholderia sp.
1.14.12.24
lipase
176% activity at 1 mg per g wet weight of cells
Burkholderia sp.
1.14.12.24
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
1.14.12.24
phosphatidylcholine
125% activity at 0.05 mg
Burkholderia sp.
1.14.12.24
phosphatidylethanolamine
144% activity at 0.1 mg per g wet weight of cells
Burkholderia sp.
1.14.12.24
Phospholipase A
125% activity at 0.01 mg
Burkholderia sp.
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.12.24
expressed in Escherichia coli PFJS39 cells
Burkholderia sp.
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.12.24
FAD
-
Burkholderia sp.
1.14.12.24
NAD+
2fold enhancement of activity in the presence of NAD+
Burkholderia sp.
1.14.12.24
NADH
5fold enhancement of activity in the presence of NADH
Burkholderia sp.
1.14.12.24
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.12.24
deoxycholate
about 68% residual activity at 6% (v/v)
Burkholderia sp.
1.14.12.24
Dodecyl sulfate
about 40% residual activity at 1% (v/v)
Burkholderia sp.
1.14.12.24
FAD
-
Burkholderia sp.
1.14.12.24
spermidine
80% residual activity at 0.05 mg
Burkholderia sp.
1.14.12.24
Triton X-100
about 50% residual activity at 2% (v/v)
Burkholderia sp.
1.14.12.24
Tween 20
about 36% residual activity at 2% (v/v)
Burkholderia sp.
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.12.24
0.05
-
O2
at pH 7.0 and 37°C
Burkholderia sp.
1.14.12.24
0.18
-
2,4-dinitrotoluene
at pH 7.0 and 37°C
Burkholderia sp.
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.12.24
Fe2+
required for activity, 238.5% activity at 1 mM
Burkholderia sp.
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.12.24
100000
-
gel filtration
Burkholderia sp.
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp.
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp. DNT
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.12.24
ammonium sulfate precipitation, column chromatography, and Sephadex G-100 gel filtration
Burkholderia sp.
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.14.12.24
2.4
-
crude extract, at pH 7.0 and 37°C
Burkholderia sp.
1.14.12.24
37.3
-
after 15.7fold, at pH 7.0 and 37°C
Burkholderia sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp.
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
1.14.12.24
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp. DNT
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.12.24
heterotrimer
1 * 52000-71000, 1 * 23000-25500, 1 * 12000-16500, SDS-PAGE
Burkholderia sp.
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.12.24
37
-
-
Burkholderia sp.
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
1.14.12.24
4
45
activity rapidly diminishes when the enzyme is heated above 45°C. At 20°C, the activity is decreased about 25% compared with 37°C. At 15°C, the activity decreases about 85% compared with 37°C, and at 4-5°C, the activity is totally absent
Burkholderia sp.
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.14.12.24
-
4
the enzyme shows 5% to 15% loss in catalytic activity when left at 0-4°C for several weeks
Burkholderia sp.
1.14.12.24
25
37
the enzyme is stable for 2 days when incubated with substrate at 37°C, 30°C, and 25°C
Burkholderia sp.
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.12.24
7
-
-
Burkholderia sp.
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.14.12.24
4
10
the enzyme is stable when incubated at pH 6 and 7, but its optimal activity is at pH 7.0. The activity is decreased significantly when incubated at pH 4.0, 5.0, 8.0, and 10.0
Burkholderia sp.