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Literature summary extracted from
- 4-Sulphobenzoate 3,4-dioxygenase. Purification and properties of a desulphonative two-component enzyme system from Comamonas testosteroni T-2 (1991), Biochem. J., 274, 833-842.
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 1.14.12.8 | ammonium sulfate precipitation leads to excessive loss of activity | Comamonas testosteroni |
| 1.14.12.8 | chloride-buffers result in diminished yields during purification | Comamonas testosteroni |
| 1.14.12.8 | glycerol, 30% v/v, stabilizes | Comamonas testosteroni |
| 1.14.12.8 | repeated freezing and thawing inactivates | Comamonas testosteroni |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.12.8 | 0.025 | 0.032 | 4-Sulfobenzoate | - |
Comamonas testosteroni |  |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.14.12.8 | cytoplasm | - |
Comamonas testosteroni | 5737 | - |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.8 | Fe2+ | requirement, iron-sulfur protein, 1 mol (2Fe-2S) per mol reductase B, one 2Fe-2S center (Rieske-type) per oxygenase A subunit, atomic absorption spectroscopy | Comamonas testosteroni | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.8 | additional information | - |
simple multi-component dioxygenase of two, perhaps three components: reductase B coupled to dimeric oxygenase, reductase C exhibits only 15% of total activity | Comamonas testosteroni |
| 1.14.12.8 | 36000 | - |
1 * 36000, reductase B, SDS-PAGE | Comamonas testosteroni |
| 1.14.12.8 | 39000 | - |
reductase B, contains 1 mol of FMN and about 2 mol each of iron and inorganic sulfur per mol, Superose gel filtration | Comamonas testosteroni |
| 1.14.12.8 | 47000 | - |
reductase B, Sephadex G-2000 gel filtration | Comamonas testosteroni |
| 1.14.12.8 | 50000 | - |
2 * 50000, oxygenase A, SDS-PAGE | Comamonas testosteroni |
| 1.14.12.8 | 85000 | - |
oxygenase A, native dioxygenase: the enzyme is presumed to be homodimeric, Superose gel filtration | Comamonas testosteroni |
| 1.14.12.8 | 105000 | - |
oxygenase A, native dioxygenase: the enzyme is presumed to be homodimeric, Sephadex G-2000 gel filtration | Comamonas testosteroni |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | Comamonas testosteroni | reaction in toluene-p-sulfonate degradation pathway, simple multi-component dioxygenase of 2, perhaps 3 components: reductase B coupled to dimeric oxygenase, the third component, reductase C, represents only 15% of total activity | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | Comamonas testosteroni T-2 | reaction in toluene-p-sulfonate degradation pathway, simple multi-component dioxygenase of 2, perhaps 3 components: reductase B coupled to dimeric oxygenase, the third component, reductase C, represents only 15% of total activity | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.12.8 | Comamonas testosteroni | - |
T-2 | - |
| 1.14.12.8 | Comamonas testosteroni T-2 | - |
T-2 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.12.8 | - |
Comamonas testosteroni |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.8 | 0.11 | - |
reductase B | Comamonas testosteroni |
| 1.14.12.8 | 0.8 | - |
oxygenase A | Comamonas testosteroni |
| 1.14.12.8 | 84 | - |
reductase B, dichlorophenolindophenol | Comamonas testosteroni |
| 1.14.12.8 | 138 | - |
reductase B, cytochrome c | Comamonas testosteroni |
| 1.14.12.8 | 318 | - |
reductase B, ferricyanide | Comamonas testosteroni |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | - |
Comamonas testosteroni | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | reaction in toluene-p-sulfonate degradation pathway, simple multi-component dioxygenase of 2, perhaps 3 components: reductase B coupled to dimeric oxygenase, the third component, reductase C, represents only 15% of total activity | Comamonas testosteroni | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | - |
Comamonas testosteroni T-2 | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
| 1.14.12.8 | 4-sulfobenzoate + NADH + O2 | reaction in toluene-p-sulfonate degradation pathway, simple multi-component dioxygenase of 2, perhaps 3 components: reductase B coupled to dimeric oxygenase, the third component, reductase C, represents only 15% of total activity | Comamonas testosteroni T-2 | 3,4-dihydroxybenzoate + sulfite + NAD+ | - |
? | |
| 1.14.12.8 | additional information | poor substrates: 2-sulfobenzoate, 3-sulfobenzoate, benzenesulfonate, 4-methylbenzenesulfonate, 4-hydroxybenzenesulfonate, 4-aminobenzenesulfonate, 4-nitrobenzenesulfonate, 4-chloro-benzenesulfonate, 4-sulfophenylacetate, 4-sulfophenylpropionate, 4-sulfophenylbutyrate, no substrates are: benzoate, 4-methylbenzoate, 4-hydroxybenzoate, 4-aminobenzoate, 4-nitrobenzoate, 4-chlorobenzoate | Comamonas testosteroni | ? | - |
? | |
| 1.14.12.8 | additional information | poor substrates: 2-sulfobenzoate, 3-sulfobenzoate, benzenesulfonate, 4-methylbenzenesulfonate, 4-hydroxybenzenesulfonate, 4-aminobenzenesulfonate, 4-nitrobenzenesulfonate, 4-chloro-benzenesulfonate, 4-sulfophenylacetate, 4-sulfophenylpropionate, 4-sulfophenylbutyrate, no substrates are: benzoate, 4-methylbenzoate, 4-hydroxybenzoate, 4-aminobenzoate, 4-nitrobenzoate, 4-chlorobenzoate | Comamonas testosteroni T-2 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.12.8 | dimer | 2 * 50000, oxygenase A, SDS-PAGE | Comamonas testosteroni |
| 1.14.12.8 | monomer | 1 * 36000, reductase B, SDS-PAGE | Comamonas testosteroni |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.8 | 25 | - |
assay at, reductase B | Comamonas testosteroni |
| 1.14.12.8 | 30 | - |
assay at | Comamonas testosteroni |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.8 | 6 | - |
assay at | Comamonas testosteroni |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.8 | FMN | flavoprotein, 1 mol FMN/mol reductase B | Comamonas testosteroni | |
| 1.14.12.8 | additional information | no independent ferredoxin | Comamonas testosteroni | |
| 1.14.12.8 | NADH | - |
Comamonas testosteroni | |
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