BRENDA - Enzyme Database show
show all sequences of 1.14.12.24

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
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
required for maximal activity, 292.3% activity at 1% (v/v)
Burkholderia sp.
ATP
124% activity at 0.1 mM
Burkholderia sp.
FMN
230.7% activity at 0.1 mM
Burkholderia sp.
GTP
116% activity at 0.1 mM
Burkholderia sp.
lipase
176% activity at 1 mg per g wet weight of cells
Burkholderia sp.
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
phosphatidylcholine
125% activity at 0.05 mg
Burkholderia sp.
phosphatidylethanolamine
144% activity at 0.1 mg per g wet weight of cells
Burkholderia sp.
Phospholipase A
125% activity at 0.01 mg
Burkholderia sp.
Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli PFJS39 cells
Burkholderia sp.
Inhibitors
Inhibitors
Commentary
Organism
Structure
deoxycholate
about 68% residual activity at 6% (v/v)
Burkholderia sp.
Dodecyl sulfate
about 40% residual activity at 1% (v/v)
Burkholderia sp.
FAD
-
Burkholderia sp.
spermidine
80% residual activity at 0.05 mg
Burkholderia sp.
Triton X-100
about 50% residual activity at 2% (v/v)
Burkholderia sp.
Tween 20
about 36% residual activity at 2% (v/v)
Burkholderia sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.05
-
O2
at pH 7.0 and 37°C
Burkholderia sp.
0.18
-
2,4-dinitrotoluene
at pH 7.0 and 37°C
Burkholderia sp.
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
required for activity, 238.5% activity at 1 mM
Burkholderia sp.
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
100000
-
gel filtration
Burkholderia sp.
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp.
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp. DNT
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Burkholderia sp.
-
-
-
Burkholderia sp. DNT
-
-
-
Purification (Commentary)
Commentary
Organism
ammonium sulfate precipitation, column chromatography, and Sephadex G-100 gel filtration
Burkholderia sp.
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4
-
crude extract, at pH 7.0 and 37°C
Burkholderia sp.
37.3
-
after 15.7fold, at pH 7.0 and 37°C
Burkholderia sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp.
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp. DNT
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
Subunits
Subunits
Commentary
Organism
heterotrimer
1 * 52000-71000, 1 * 23000-25500, 1 * 12000-16500, SDS-PAGE
Burkholderia sp.
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
-
Burkholderia sp.
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
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]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
-
4
the enzyme shows 5% to 15% loss in catalytic activity when left at 0-4°C for several weeks
Burkholderia sp.
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
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
-
Burkholderia sp.
pH Range
pH Minimum
pH Maximum
Commentary
Organism
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
Cofactor
Commentary
Organism
Structure
FAD
-
Burkholderia sp.
NAD+
2fold enhancement of activity in the presence of NAD+
Burkholderia sp.
NADH
5fold enhancement of activity in the presence of NADH
Burkholderia sp.
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
required for maximal activity, 292.3% activity at 1% (v/v)
Burkholderia sp.
ATP
124% activity at 0.1 mM
Burkholderia sp.
FMN
230.7% activity at 0.1 mM
Burkholderia sp.
GTP
116% activity at 0.1 mM
Burkholderia sp.
lipase
176% activity at 1 mg per g wet weight of cells
Burkholderia sp.
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
phosphatidylcholine
125% activity at 0.05 mg
Burkholderia sp.
phosphatidylethanolamine
144% activity at 0.1 mg per g wet weight of cells
Burkholderia sp.
Phospholipase A
125% activity at 0.01 mg
Burkholderia sp.
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli PFJS39 cells
Burkholderia sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
-
Burkholderia sp.
NAD+
2fold enhancement of activity in the presence of NAD+
Burkholderia sp.
NADH
5fold enhancement of activity in the presence of NADH
Burkholderia sp.
NADP+
2fold enhancement of activity in the presence of NADP+
Burkholderia sp.
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
deoxycholate
about 68% residual activity at 6% (v/v)
Burkholderia sp.
Dodecyl sulfate
about 40% residual activity at 1% (v/v)
Burkholderia sp.
FAD
-
Burkholderia sp.
spermidine
80% residual activity at 0.05 mg
Burkholderia sp.
Triton X-100
about 50% residual activity at 2% (v/v)
Burkholderia sp.
Tween 20
about 36% residual activity at 2% (v/v)
Burkholderia sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.05
-
O2
at pH 7.0 and 37°C
Burkholderia sp.
0.18
-
2,4-dinitrotoluene
at pH 7.0 and 37°C
Burkholderia sp.
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
required for activity, 238.5% activity at 1 mM
Burkholderia sp.
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
100000
-
gel filtration
Burkholderia sp.
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp.
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
2,4-dinitrotoluene + NADH + H+ + O2
Burkholderia sp. DNT
-
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
ammonium sulfate precipitation, column chromatography, and Sephadex G-100 gel filtration
Burkholderia sp.
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4
-
crude extract, at pH 7.0 and 37°C
Burkholderia sp.
37.3
-
after 15.7fold, at pH 7.0 and 37°C
Burkholderia sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp.
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
2,4-dinitrotoluene + NADH + H+ + O2
-
733523
Burkholderia sp. DNT
4-methyl-5-nitrocatechol + nitrite + NAD+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
heterotrimer
1 * 52000-71000, 1 * 23000-25500, 1 * 12000-16500, SDS-PAGE
Burkholderia sp.
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
-
Burkholderia sp.
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
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)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
-
4
the enzyme shows 5% to 15% loss in catalytic activity when left at 0-4°C for several weeks
Burkholderia sp.
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)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
-
Burkholderia sp.
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
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.
Other publictions for EC 1.14.12.24
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
741610
Kuece
-
Biodegradation of 2,4-dinitro ...
Microbacterium saccharophilum
Ann. Microbiol.
65
467-476
2015
-
-
-
-
-
-
-
-
-
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1
-
1
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1
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1
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1
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1
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1
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-
-
-
-
-
-
-
733523
Khleifat
-
Characterization of 2,4-dinitr ...
Burkholderia sp., Burkholderia sp. DNT
Bioremediat. J.
14
38-53
2010
9
-
1
-
-
-
6
2
-
1
1
2
-
8
-
-
1
-
-
-
2
-
2
1
1
1
2
-
1
1
-
4
-
-
-
9
-
1
4
-
-
-
-
6
-
2
-
1
1
2
-
-
-
1
-
-
2
-
2
1
1
1
2
-
1
1
-
-
-
-
-
-
-
-
733549
Leungsakul T
Oxidation of aminonitrotoluene ...
Burkholderia sp., Burkholderia sp. DNT
Biotechnol. Bioeng.
93
231-237
2006
-
-
1
-
1
-
-
-
-
-
-
2
-
16
-
-
-
-
-
-
-
-
27
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
27
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733548
Leungsakul
Saturation mutagenesis of 2,4- ...
Burkholderia sp., Burkholderia sp. DNT
Biotechnol. Bioeng.
92
416-426
2005
-
-
1
-
2
-
-
-
-
-
-
4
-
5
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733152
Keenan
Saturation mutagenesis of Burk ...
Burkholderia cepacia, Burkholderia cepacia R34, Burkholderia sp., Burkholderia sp. DNT
Appl. Environ. Microbiol.
70
3222-3231
2004
-
-
1
-
4
-
-
-
-
-
-
2
-
9
-
-
-
-
-
-
-
-
24
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
24
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
734412
Lin
Effects of Vitreoscilla hemogl ...
Burkholderia sp., Burkholderia sp. DNT
J. Ind. Microbiol. Biotechnol.
30
362-368
2003
1
-
-
-
-
-
-
1
1
-
2
2
-
11
-
-
1
-
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
1
-
2
2
-
-
-
1
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
734502
Fish P.A
-
Vitreoscilla hemoglobin enhanc ...
Burkholderia sp., Burkholderia sp. DNT
J. Mol. Catal. B
9
75-82
2000
1
-
1
-
-
-
-
-
-
-
-
2
-
8
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
734057
Parales
Enzyme specificity of 2-nitrot ...
Burkholderia sp., Burkholderia sp. DNT
J. Bacteriol.
180
1194-1199
1998
-
-
-
-
1
-
-
-
-
-
-
-
-
11
-
-
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-
-
-
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-
8
-
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1
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-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
734058
Parales
Substrate specificities of hyb ...
Burkholderia sp., Burkholderia sp. DNT
J. Bacteriol.
180
2337-2344
1998
-
-
1
-
1
-
-
-
-
-
1
2
-
14
-
-
-
-
-
-
1
-
10
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
-
1
2
-
-
-
-
-
-
1
-
10
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
734055
Suen
2,4-Dinitrotoluene dioxygenase ...
Burkholderia sp., Burkholderia sp. DNT
J. Bacteriol.
178
4926-4934
1996
-
-
1
-
-
-
-
-
-
-
-
2
-
16
-
-
-
-
-
-
-
-
25
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
25
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
734052
Suen
Cloning and characterization o ...
Pseudomonas sp., Pseudomonas sp. DNT
J. Bacteriol.
175
1831-1837
1993
-
-
-
-
-
-
-
-
-
-
-
2
-
5
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
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-
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-
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-
2
-
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-
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-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733145
Spanggord
Biodegradation of 2,4-dinitrot ...
Pseudomonas sp., Pseudomonas sp. DNT
Appl. Environ. Microbiol.
57
3200-3205
1991
-
-
-
-
-
-
-
-
-
-
-
2
-
4
-
-
-
-
-
-
-
-
12
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
12
-
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-
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