BRENDA - Enzyme Database show

4-Sulphobenzoate 3,4-dioxygenase. Purification and properties of a desulphonative two-component enzyme system from Comamonas testosteroni T-2

Locher, H.H.; Leisinger, T.; Cook, A.M.; Biochem. J. 274, 833-842 (1991)

Data extracted from this reference:

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
Commentary
Organism
Structure
1.14.12.8
0.025
0.032
4-Sulfobenzoate
-
Comamonas testosteroni
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.14.12.8
cytoplasm
-
Comamonas testosteroni
5737
-
Metals/Ions
EC Number
Metals/Ions
Commentary
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]
Commentary
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
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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+
-
Comamonas testosteroni
?
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+
-
Comamonas testosteroni T-2
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.12.8
Comamonas testosteroni
-
T-2
-
1.14.12.8
Comamonas testosteroni T-2
-
T-2
-
Purification (Commentary)
EC Number
Commentary
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]
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.8
4-sulfobenzoate + NADH + O2
-
439044
Comamonas testosteroni
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni
?
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
439044
Comamonas testosteroni
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni
?
1.14.12.8
4-sulfobenzoate + NADH + O2
-
439044
Comamonas testosteroni T-2
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni T-2
?
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
439044
Comamonas testosteroni T-2
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni T-2
?
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
439044
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
439044
Comamonas testosteroni T-2
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
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]
Commentary
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
Commentary
Organism
1.14.12.8
6
-
assay at
Comamonas testosteroni
Cofactor
EC Number
Cofactor
Commentary
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
Cofactor (protein specific)
EC Number
Cofactor
Commentary
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
General Stability (protein specific)
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.12.8
0.025
0.032
4-Sulfobenzoate
-
Comamonas testosteroni
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.14.12.8
cytoplasm
-
Comamonas testosteroni
5737
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
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] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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+
-
Comamonas testosteroni
?
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+
-
Comamonas testosteroni T-2
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.12.8
-
Comamonas testosteroni
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.14.12.8
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) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.12.8
4-sulfobenzoate + NADH + O2
-
439044
Comamonas testosteroni
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni
?
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
439044
Comamonas testosteroni
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni
?
1.14.12.8
4-sulfobenzoate + NADH + O2
-
439044
Comamonas testosteroni T-2
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni T-2
?
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
439044
Comamonas testosteroni T-2
3,4-dihydroxybenzoate + sulfite + NAD+
-
439044
Comamonas testosteroni T-2
?
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
439044
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
439044
Comamonas testosteroni T-2
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
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] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.12.8
25
-
assay at, reductase B
Comamonas testosteroni
1.14.12.8
30
-
assay at
Comamonas testosteroni
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.12.8
6
-
assay at
Comamonas testosteroni