BRENDA - Enzyme Database show

Purification and properties of a plasmid-encoded 2,4-dichlorophenol hydroxylase

Liu, T.; Chapman, P.J.; FEBS Lett. 173, 314-318 (1984)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.13.20
expression in Pseudomonas putida
Cupriavidus necator
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.13.20
2,4-Dichlorophenol
above 1 mM
Cupriavidus necator
1.14.13.20
AgNO3
-
Cupriavidus necator
1.14.13.20
CuSO4
-
Cupriavidus necator
1.14.13.20
diethyldithiocarbamate
-
Cupriavidus necator
1.14.13.20
Dithionitrobenzoate
-
Cupriavidus necator
1.14.13.20
dithiothreitol
slight
Cupriavidus necator
1.14.13.20
FeSO4
-
Cupriavidus necator
1.14.13.20
Hg2+
HgCl2
Cupriavidus necator
1.14.13.20
KCl
high salt concentration inhibits, 0.05 M: 40%, 0.1 M: 50%, 0.2 M: 70%
Cupriavidus necator
1.14.13.20
KCN
no effect
Cupriavidus necator
1.14.13.20
o-phenanthroline
-
Cupriavidus necator
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.20
0.002
-
2,4-Dichlorophenol
cosubstrate NADPH
Cupriavidus necator
1.14.13.20
0.004
-
2,4-Dichlorophenol
cosubstrate NADH
Cupriavidus necator
1.14.13.20
0.048
-
NADPH
+ 2,4-dichlorophenol
Cupriavidus necator
1.14.13.20
0.3
-
NADH
+ 2,4-dichlorophenol
Cupriavidus necator
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.13.20
45000
-
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
1.14.13.20
67000
-
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
1.14.13.20
224000
-
ultracentrifugation
Cupriavidus necator
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.13.20
2,4-dichlorophenol + NADPH + O2
Cupriavidus necator
-
3,5-dichlorocatechol + NADP+ + H2O
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
Cupriavidus necator JMP 134-1
-
3,5-dichlorocatechol + NADP+ + H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.13.20
Cupriavidus necator
-
basonym Alcaligenes eutrophus; JMP134
-
1.14.13.20
Cupriavidus necator JMP 134-1
-
JMP134
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.13.20
plasmid pJP4-encoded enzyme from Alcaligenes eutrophus JMP134 transferred to Pseudomonas putida 3CB5 and Alcaligenes eutrophus 335
Cupriavidus necator
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.14.13.20
1.48
-
-
Cupriavidus necator
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.20
2,4,5-trichlorophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2,4,5-trichlorophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
2,4-dibromophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2,4-dibromophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
-
439012
Cupriavidus necator
3,5-dichlorocatechol + NADP+ + H2O
-
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
3,5-dichlorocatechol + NADP+ + H2O
-
-
-
?
1.14.13.20
2-chloro-4-nitrophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2-chlorophenol + NADPH + O2
incompletely metabolized
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2-chlorophenol + NADPH + O2
incompletely metabolized
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
4-chloro-2-methylphenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
4-chloro-2-methylphenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
4-chloro-2-nitrophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
4-chlorophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.13.20
tetramer
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.13.20
7.3
8
phosphate buffer
Cupriavidus necator
1.14.13.20
8.1
8.5
100 mM Tris-HCl buffer
Cupriavidus necator
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.20
FAD
1 mol of enzyme contains: 3.5 mol FAD; flavoprotein
Cupriavidus necator
1.14.13.20
NADH
50% of the activity with NADPH
Cupriavidus necator
1.14.13.20
NADPH
-
Cupriavidus necator
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.13.20
expression in Pseudomonas putida
Cupriavidus necator
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.20
FAD
1 mol of enzyme contains: 3.5 mol FAD; flavoprotein
Cupriavidus necator
1.14.13.20
NADH
50% of the activity with NADPH
Cupriavidus necator
1.14.13.20
NADPH
-
Cupriavidus necator
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.13.20
2,4-Dichlorophenol
above 1 mM
Cupriavidus necator
1.14.13.20
AgNO3
-
Cupriavidus necator
1.14.13.20
CuSO4
-
Cupriavidus necator
1.14.13.20
diethyldithiocarbamate
-
Cupriavidus necator
1.14.13.20
Dithionitrobenzoate
-
Cupriavidus necator
1.14.13.20
dithiothreitol
slight
Cupriavidus necator
1.14.13.20
FeSO4
-
Cupriavidus necator
1.14.13.20
Hg2+
HgCl2
Cupriavidus necator
1.14.13.20
KCl
high salt concentration inhibits, 0.05 M: 40%, 0.1 M: 50%, 0.2 M: 70%
Cupriavidus necator
1.14.13.20
KCN
no effect
Cupriavidus necator
1.14.13.20
o-phenanthroline
-
Cupriavidus necator
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.20
0.002
-
2,4-Dichlorophenol
cosubstrate NADPH
Cupriavidus necator
1.14.13.20
0.004
-
2,4-Dichlorophenol
cosubstrate NADH
Cupriavidus necator
1.14.13.20
0.048
-
NADPH
+ 2,4-dichlorophenol
Cupriavidus necator
1.14.13.20
0.3
-
NADH
+ 2,4-dichlorophenol
Cupriavidus necator
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.13.20
45000
-
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
1.14.13.20
67000
-
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
1.14.13.20
224000
-
ultracentrifugation
Cupriavidus necator
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.13.20
2,4-dichlorophenol + NADPH + O2
Cupriavidus necator
-
3,5-dichlorocatechol + NADP+ + H2O
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
Cupriavidus necator JMP 134-1
-
3,5-dichlorocatechol + NADP+ + H2O
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.13.20
plasmid pJP4-encoded enzyme from Alcaligenes eutrophus JMP134 transferred to Pseudomonas putida 3CB5 and Alcaligenes eutrophus 335
Cupriavidus necator
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.14.13.20
1.48
-
-
Cupriavidus necator
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.20
2,4,5-trichlorophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2,4,5-trichlorophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
2,4-dibromophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2,4-dibromophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
-
439012
Cupriavidus necator
3,5-dichlorocatechol + NADP+ + H2O
-
-
-
?
1.14.13.20
2,4-dichlorophenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
3,5-dichlorocatechol + NADP+ + H2O
-
-
-
?
1.14.13.20
2-chloro-4-nitrophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2-chlorophenol + NADPH + O2
incompletely metabolized
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
2-chlorophenol + NADPH + O2
incompletely metabolized
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
4-chloro-2-methylphenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
4-chloro-2-methylphenol + NADPH + O2
-
439012
Cupriavidus necator JMP 134-1
?
-
-
-
?
1.14.13.20
4-chloro-2-nitrophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
1.14.13.20
4-chlorophenol + NADPH + O2
-
439012
Cupriavidus necator
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.13.20
tetramer
alpha2beta2, 2 * 67000 + 2 * 45000, SDS-PAGE
Cupriavidus necator
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.13.20
7.3
8
phosphate buffer
Cupriavidus necator
1.14.13.20
8.1
8.5
100 mM Tris-HCl buffer
Cupriavidus necator