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Literature summary extracted from
- Purification and some properties of 2-halobenzoate 1,2-dioxygenase, a two-component enzyme system from Pseudomonas cepacia 2CBS (1992), J. Bacteriol., 174, 279-290.
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.13 | 2,2'-dipyridyl | 100% inhibition at 1 mM | Burkholderia cepacia |  |
| 1.14.12.13 | Cu2+ | 100% inhibition compared to the activity without any metal | Burkholderia cepacia | |
| 1.14.12.13 | EDTA | 100% inhibition at 1 mM | Burkholderia cepacia | |
| 1.14.12.13 | iodoacetate | 55% inhibition at 2 mM | Burkholderia cepacia | |
| 1.14.12.13 | KCN | 40% inhibition at 2 mM | Burkholderia cepacia | |
| 1.14.12.13 | additional information | no inhibition with sodium azide | Burkholderia cepacia | |
| 1.14.12.13 | N-ethylmaleimide | 100% inhibition at 2 mM | Burkholderia cepacia | |
| 1.14.12.13 | Ni2+ | 25% inhibition compared to the activity without any metal | Burkholderia cepacia | |
| 1.14.12.13 | o-phenanthroline | 100% inhibition at 0.5 mM | Burkholderia cepacia | |
| 1.14.12.13 | p-chloromercuribenzoate | 17% inhibition at 0.005 mM | Burkholderia cepacia | |
| 1.14.12.13 | Zn2+ | 40% inhibition compared to the activity without any metal | Burkholderia cepacia | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.12.13 | 0.023 | - |
2-Chlorobenzoate | component A | Burkholderia cepacia | |
| 1.14.12.13 | 0.079 | - |
NADH | component B | Burkholderia cepacia | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.13 | Fe2+ | exogenous, required for activity | Burkholderia cepacia | |
| 1.14.12.13 | Fe2+ | component B contains 1.7 mol and A 5.8 mol of iron per mol of enzyme | Burkholderia cepacia | |
| 1.14.12.13 | Sulfide | component B contains 1.7 mol and A 6.0 mol of acid-labile sulfide per mol of enzyme | Burkholderia cepacia | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.13 | 20000 | - |
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE | Burkholderia cepacia |
| 1.14.12.13 | 37500 | - |
1 * 37500, component B, SDS-PAGE | Burkholderia cepacia |
| 1.14.12.13 | 37500 | 38000 | component B, gel filtration | Burkholderia cepacia |
| 1.14.12.13 | 52000 | - |
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE | Burkholderia cepacia |
| 1.14.12.13 | 200000 | 220000 | component A, gel filtration | Burkholderia cepacia |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.12.13 | 2-chlorobenzoate + NADH + H+ + O2 | Burkholderia cepacia | first step of 2-chlorobenzoate degradation via meta-cleavage | catechol + chloride + NAD+ + CO2 | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.12.13 | Burkholderia cepacia | - |
2CBS, isolated from water | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.12.13 | component A: anion-exchange, gel filtration, component B: anion-exchange, affinity and adsorption chromatography | Burkholderia cepacia |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.14.12.13 | 2-chlorobenzoate + NADH + H+ + O2 = catechol + chloride + NAD+ + CO2 | inducible two-component enzyme system: component B is an iron-sulfur flavoprotein (reductase) and red-brown component A (oxygenase) is an iron-sulfur protein | Burkholderia cepacia | |
| 1.14.12.13 | 2-chlorobenzoate + NADH + H+ + O2 = catechol + chloride + NAD+ + CO2 | multi-component dioxygenase system proposed: 1,2-hydroxylating, dehalogenating, decarboxylating | Burkholderia cepacia |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.13 | 0.0085 | - |
crude extract, 3.4 mg protein per ml in the assay | Burkholderia cepacia |
| 1.14.12.13 | 0.53 | - |
activity of component A after gel filtration in presence of component B | Burkholderia cepacia |
| 1.14.12.13 | 4 | - |
activity of component B in presence of component A | Burkholderia cepacia |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.12.13 | -20°C component A, 80-85% loss of activity | Burkholderia cepacia |
| 1.14.12.13 | -20°C, component A, 20% glycerol, 50% loss of activity | Burkholderia cepacia |
| 1.14.12.13 | -20°C, component B, 20% sucrose, 2 weeks, 25% loss of activity | Burkholderia cepacia |
| 1.14.12.13 | -80°C or -20°C, component B, 2 weeks, 70% loss of activity | Burkholderia cepacia |
| 1.14.12.13 | -80°C or -20°C, component B, 20% glycerol, 2 weeks, 15% loss of activity | Burkholderia cepacia |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.12.13 | 2-aminobenzoate + NADH + O2 | - |
Burkholderia cepacia | ? + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-bromobenzoate + NADH + O2 | - |
Burkholderia cepacia | catechol + bromide + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-chlorobenzoate + NADH + H+ + O2 | - |
Burkholderia cepacia | catechol + chloride + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-chlorobenzoate + NADH + H+ + O2 | first step of 2-chlorobenzoate degradation via meta-cleavage | Burkholderia cepacia | catechol + chloride + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-fluorobenzoate + NADH + O2 | - |
Burkholderia cepacia | catechol + fluoride + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-hydroxybenzoate + NADH + O2 | - |
Burkholderia cepacia | catechol + ? + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-iodobenzoate + NADH + O2 | - |
Burkholderia cepacia | catechol + iodide + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-methoxybenzoate + NADH + O2 | - |
Burkholderia cepacia | catechol + ? + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | 2-methylbenzoate + NADH + O2 | - |
Burkholderia cepacia | o-cresol + ? + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | anthranilate + NADH + O2 | - |
Burkholderia cepacia | catechol + ? + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | benzoate + NADH + H+ + O2 | - |
Burkholderia cepacia | catechol + NAD+ + CO2 | - |
? | |
| 1.14.12.13 | additional information | very broad substrate specificity is listed, benzoate analogs with substituents in ortho position are preferred. Trichloroethylene, dihydroxybenzoates, benzene chlorobenzene, toluene, phenol and chlorophenylacetate isomers do not serve as substrates | Burkholderia cepacia | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.12.13 | hexamer | 3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE | Burkholderia cepacia |
| 1.14.12.13 | monomer | 1 * 37500, component B, SDS-PAGE | Burkholderia cepacia |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.13 | 25 | - |
assay at | Burkholderia cepacia |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.13 | 6.5 | - |
assay at | Burkholderia cepacia |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.13 | FAD | component B contains 0.8 mol per mol of enzyme, 0.002 mM increases activity | Burkholderia cepacia | |
| 1.14.12.13 | FMN | little increase of activity by addition of 0.002 mM | Burkholderia cepacia | |
| 1.14.12.13 | NADH | can not be replaced by NADPH | Burkholderia cepacia | |
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