BRENDA - Enzyme Database

Histidine 90 function in 4-chlorobenzoyl-coenzyme A dehalogenase catalysis

Zhang, W.; Wei, Y.; Luo, L.; Taylor, K.L.; Yang, G.; Dunaway-Mariano, D.; Benning, M.M.; Holden, H.M.; Biochemistry 40, 13474-13482 (2001)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
3.8.1.7
purified recombinant mutant H90Q complexed with the substrate, 8% PEG 8000, 0.2 M potassium chloride, 50 mM CHES, pH 9.0, 5 mM NaN3, 4 °C, X-ray diffraction structure determination and analysis at 1.8 A resolution
Pseudomonas sp.
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.8.1.7
H90Q
site-directed mutagenesis, exchange of the active site H90, reduced formation of arylated enzyme intermediate, and 154fold reduction of arylated enzyme intermediate hydrolysis, active site structure
Pseudomonas sp.
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.8.1.7
additional information
-
additional information
kinetics
Pseudomonas sp.
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.8.1.7
4-chlorobenzoyl-CoA + H2O
Pseudomonas sp.
-
4-hydroxybenzoyl-CoA + chloride
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.8.1.7
Pseudomonas sp.
A5JTM5
-
-
Reaction
EC Number
Reaction
Commentary
Organism
3.8.1.7
4-chlorobenzoyl-CoA + H2O = 4-hydroxybenzoyl CoA + chloride
specific for dehalogenation at the 4-position, can dehalogenate substrates bearing fluorine, chlorine, bromine and iodine in the 4-position, mechanism, attack of Asp145 on the C(4) of the substrates' benzoyl ring to form a Meisenheimer intermediate, followed by expulsion of chloride ion to form an arylated enzyme intermediate, and ester hydrolysis to form 4-hydroxybenzoyl-CoA, active site residue His90 is involved and important for the regeneration of the active enzyme
Pseudomonas sp.
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.8.1.7
1.5
-
purified wild-type enzyme
Pseudomonas sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.8.1.7
4-chlorobenzoyl-CoA + H2O
-
654585
Pseudomonas sp.
4-hydroxybenzoyl-CoA + chloride
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.8.1.7
25
-
assay at
Pseudomonas sp.
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.8.1.7
additional information
-
additional information
turnover rates for dormations of intermediates, wildtype and mutant
Pseudomonas sp.
3.8.1.7
0.0002
-
4-chlorobenzoyl-CoA
multiple turnovers, mutant H90Q, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
0.016
-
4-chlorobenzoyl-CoA
single turnover, mutant H90Q, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
0.7
-
4-chlorobenzoyl-CoA
multiple turnovers, wild-type enzyme, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
2.3
-
4-chlorobenzoyl-CoA
single turnover, wild-type enzyme, pH 7.5, 25°C
Pseudomonas sp.
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.8.1.7
7.5
-
assay at
Pseudomonas sp.
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
3.8.1.7
purified recombinant mutant H90Q complexed with the substrate, 8% PEG 8000, 0.2 M potassium chloride, 50 mM CHES, pH 9.0, 5 mM NaN3, 4 °C, X-ray diffraction structure determination and analysis at 1.8 A resolution
Pseudomonas sp.
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.8.1.7
H90Q
site-directed mutagenesis, exchange of the active site H90, reduced formation of arylated enzyme intermediate, and 154fold reduction of arylated enzyme intermediate hydrolysis, active site structure
Pseudomonas sp.
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.8.1.7
additional information
-
additional information
kinetics
Pseudomonas sp.
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.8.1.7
4-chlorobenzoyl-CoA + H2O
Pseudomonas sp.
-
4-hydroxybenzoyl-CoA + chloride
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.8.1.7
1.5
-
purified wild-type enzyme
Pseudomonas sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.8.1.7
4-chlorobenzoyl-CoA + H2O
-
654585
Pseudomonas sp.
4-hydroxybenzoyl-CoA + chloride
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.8.1.7
25
-
assay at
Pseudomonas sp.
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.8.1.7
additional information
-
additional information
turnover rates for dormations of intermediates, wildtype and mutant
Pseudomonas sp.
3.8.1.7
0.0002
-
4-chlorobenzoyl-CoA
multiple turnovers, mutant H90Q, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
0.016
-
4-chlorobenzoyl-CoA
single turnover, mutant H90Q, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
0.7
-
4-chlorobenzoyl-CoA
multiple turnovers, wild-type enzyme, pH 7.5, 25°C
Pseudomonas sp.
3.8.1.7
2.3
-
4-chlorobenzoyl-CoA
single turnover, wild-type enzyme, pH 7.5, 25°C
Pseudomonas sp.
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.8.1.7
7.5
-
assay at
Pseudomonas sp.