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Literature summary for 1.14.12.24 extracted from

  • Khleifat, K.M.
    Characterization of 2,4-dinitrotoluene dioxygenase from recombinant Escherichia coli strain PFJS39: Its direct interaction with vitreoscilla hemoglobin (2010), Bioremediat. J., 14, 38-53.
No PubMed abstract available

Activating Compound

Activating Compound Comment 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)

Cloned (Comment) Organism
expressed in Escherichia coli PFJS39 cells Burkholderia sp.

Inhibitors

Inhibitors Comment 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 Comment 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 Comment Organism Structure
Fe2+ required for activity, 238.5% activity at 1 mM Burkholderia sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
100000
-
gel filtration Burkholderia sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment Textmining
Burkholderia sp.
-
-
-
Burkholderia sp. DNT
-
-
-

Purification (Commentary)

Purification (Comment) 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] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
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+
-
?

Subunits

Subunits Comment Organism
heterotrimer 1 * 52000-71000, 1 * 23000-25500, 1 * 12000-16500, SDS-PAGE Burkholderia sp.

Synonyms

Synonyms Comment Organism
DNT dioxygenase
-
Burkholderia sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Burkholderia sp.

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment 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] Comment 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 Comment Organism
7
-
-
Burkholderia sp.

pH Range

pH Minimum pH Maximum Comment 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 Comment 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.