Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
F64A | site-directed mutagenesis, decreased kcat and slightly increased Km compared to the wild-type enzyme | Pseudomonas sp. |
F64P | site-directed mutagenesis, highly decreased kcat compared to the wild-type enzyme, nearly inactive | Pseudomonas sp. |
G113A | site-directed mutagenesis, decreased kcat and increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R24K | site-directed mutagenesis, increased kcat and increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R24L | site-directed mutagenesis, decreased kcat and increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R257K | site-directed mutagenesis, slightly decreased kcat and slightly increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R257L | site-directed mutagenesis, decreased kcat and increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R67K | site-directed mutagenesis, decreased kcat and slightly increased Km compared to the wild-type enzyme | Pseudomonas sp. |
R67L | site-directed mutagenesis, expression in inclusion bodies | Pseudomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3'-AMP | weak inhibitor | Pseudomonas sp. | |
3'-dephospho-CoA | - |
Pseudomonas sp. | |
4-Chlorobenzoate | poor inhibitor | Pseudomonas sp. | |
4-chlorobenzoyl-coenzyme A | competitive | Pseudomonas sp. | |
4-hydroxybenzoyl-3'-dephospho-CoA | competitive | Pseudomonas sp. | |
4-hydroxybenzoyl-CoA | competitive | Pseudomonas sp. | |
4-hydroxybenzoyl-dithio-CoA | competitive | Pseudomonas sp. | |
4-methylbenzoyl-3'-dephospho-CoA | competitive | Pseudomonas sp. | |
4-methylbenzoyl-CoA | competitive | Pseudomonas sp. | |
5'-ADP | weak inhibitor | Pseudomonas sp. | |
5'-AMP | weak inhibitor | Pseudomonas sp. | |
adenosine | weak inhibitor | Pseudomonas sp. | |
adenosine 3',5'-bisphosphate | weak inhibitor | Pseudomonas sp. | |
ATP | weak inhibitor | Pseudomonas sp. | |
CoA | feedback inhibition | Pseudomonas sp. | |
diphosphate | weak inhibitor | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Pseudomonas sp. | |
0.0037 | - |
4-chlorobenzoyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.0052 | - |
4-chlorobenzoyl-CoA | mutant F64A, pH 7.5, 25°C | Pseudomonas sp. | |
0.0058 | - |
4-chlorobenzoyl-CoA | mutant R257K, pH 7.5, 25°C | Pseudomonas sp. | |
0.0069 | - |
4-chlorobenzoyl-CoA | mutant R67K, pH 7.5, 25°C | Pseudomonas sp. | |
0.014 | - |
4-chlorobenzoyl-CoA | mutant G113A, pH 7.5, 25°C | Pseudomonas sp. | |
0.02 | - |
4-chlorobenzoyl-3'-dephospho-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.021 | - |
4-chlorobenzoyl-CoA | mutant R24K, pH 7.5, 25°C | Pseudomonas sp. | |
0.024 | - |
4-bromophenacyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.033 | - |
4-chlorobenzoyl-dithio-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.041 | - |
4-chlorobenzoyl-CoA | mutant R24L, pH 7.5, 25°C | Pseudomonas sp. | |
0.07 | - |
4-chlorobenzoyl-CoA | mutant R257L, pH 7.5, 25°C | Pseudomonas sp. | |
0.07 | - |
4-chlorobenzoyl-pantetheine | wild-type enzyme, pH 7.5, 25°C, in presence of 1 mM 5'-ADP | Pseudomonas sp. | |
0.11 | - |
4-chlorobenzoyl-CoA | mutant G114A, pH 7.5, 25°C | Pseudomonas sp. | |
0.31 | - |
4-chlorobenzoyl-epsilon-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.44 | - |
4-chlorobenzoyl-pantetheine phosphate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
2 | - |
4-chlorobenzoyl-pantetheine | above, wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-chlorobenzoyl-CoA + H2O | Pseudomonas sp. | degradation in soil, pathway overview | 4-hydroxybenzoyl-CoA + chloride | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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, multistep 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 structure | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-bromophenacyl-CoA + H2O | - |
Pseudomonas sp. | 4-hydroxyphenacyl-CoA + bromide | - |
? | |
4-chlorobenzoyl-3'-dephospho-CoA + H2O | - |
Pseudomonas sp. | 4-hydroxybenzoyl-3'-dephospho-CoA + chloride | - |
? | |
4-chlorobenzoyl-CoA + H2O | - |
Pseudomonas sp. | 4-hydroxybenzoyl-CoA + chloride | - |
? | |
4-chlorobenzoyl-CoA + H2O | degradation in soil, pathway overview | Pseudomonas sp. | 4-hydroxybenzoyl-CoA + chloride | - |
? | |
4-chlorobenzoyl-dithio-CoA + H2O | - |
Pseudomonas sp. | 4-hydroxybenzoyl-dithio-CoA + chloride | - |
? | |
4-chlorobenzoyl-epsilon-CoA + H2O | - |
Pseudomonas sp. | 4-hydroxybenzoyl-epsilon-CoA + chloride | - |
? | |
4-chlorobenzoyl-pantetheine + H2O | very low activity | Pseudomonas sp. | 4-hydroxybenzoyl-pantetheine + chloride | - |
? | |
4-chlorobenzoyl-pantetheine phosphate + H2O | - |
Pseudomonas sp. | 4-hydroxybenzoyl-pantetheine phosphate + chloride | - |
? |
Synonyms | Comment | Organism |
---|---|---|
4-CBA-CoA dehalogenase | - |
Pseudomonas sp. |
4-chlorobenzoyl-coenzyme A dehalogenase | - |
Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00001 | - |
4-chlorobenzoyl-CoA | below, mutant F64P, pH 7.5, 25°C | Pseudomonas sp. | |
0.001 | - |
4-chlorobenzoyl-pantetheine | about, wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.002 | - |
4-chlorobenzoyl-CoA | mutant G114A, pH 7.5, 25°C | Pseudomonas sp. | |
0.003 | - |
4-chlorobenzoyl-dithio-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.005 | - |
4-chlorobenzoyl-CoA | mutant G113A, pH 7.5, 25°C | Pseudomonas sp. | |
0.007 | - |
4-chlorobenzoyl-epsilon-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.03 | - |
4-bromophenacyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.03 | - |
4-chlorobenzoyl-CoA | mutant F64A, pH 7.5, 25°C | Pseudomonas sp. | |
0.051 | - |
4-chlorobenzoyl-pantetheine phosphate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.056 | - |
4-chlorobenzoyl-pantetheine | wild-type enzyme, pH 7.5, 25°C, in presence of 1 mM 5'-ADP | Pseudomonas sp. | |
0.14 | - |
4-chlorobenzoyl-CoA | mutant R257L, pH 7.5, 25°C | Pseudomonas sp. | |
0.33 | - |
4-chlorobenzoyl-CoA | mutant R24L, pH 7.5, 25°C | Pseudomonas sp. | |
0.38 | - |
4-chlorobenzoyl-CoA | mutant R67K, pH 7.5, 25°C | Pseudomonas sp. | |
0.45 | - |
4-chlorobenzoyl-3'-dephospho-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.55 | - |
4-chlorobenzoyl-CoA | mutant R257K, pH 7.5, 25°C | Pseudomonas sp. | |
0.6 | - |
4-chlorobenzoyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.95 | - |
4-chlorobenzoyl-CoA | mutant R24K, pH 7.5, 25°C | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0017 | - |
4-hydroxybenzoyl-CoA | mutant R257K, pH 7.5, 25°C | Pseudomonas sp. | |
0.0024 | - |
4-hydroxybenzoyl-CoA | mutant R24K, pH 7.5, 25°C | Pseudomonas sp. | |
0.0025 | - |
4-hydroxybenzoyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.0031 | - |
4-methylbenzoyl-CoA | mutant R257K, pH 7.5, 25°C | Pseudomonas sp. | |
0.0042 | - |
4-methylbenzoyl-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.0062 | - |
4-hydroxybenzoyl-CoA | mutant R67K, pH 7.5, 25°C | Pseudomonas sp. | |
0.011 | - |
4-methylbenzoyl-CoA | mutant R24K, pH 7.5, 25°C | Pseudomonas sp. | |
0.0165 | - |
4-hydroxybenzoyl-CoA | mutant R24L, pH 7.5, 25°C | Pseudomonas sp. | |
0.019 | - |
4-hydroxybenzoyl-CoA | mutant R257L, pH 7.5, 25°C | Pseudomonas sp. | |
0.022 | - |
4-methylbenzoyl-3'-dephospho-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.04 | - |
4-hydroxybenzoyl-CoA | mutant G113A, pH 7.5, 25°C | Pseudomonas sp. | |
0.044 | - |
4-methylbenzoyl-CoA | mutant G113A, pH 7.5, 25°C | Pseudomonas sp. | |
0.046 | - |
4-methylbenzoyl-CoA | mutant R24L, pH 7.5, 25°C | Pseudomonas sp. | |
0.05 | 0.1 | 4-hydroxybenzoyl-CoA | mutant G114A, pH 7.5, 25°C | Pseudomonas sp. | |
0.051 | - |
4-methylbenzoyl-CoA | mutant R67K, pH 7.5, 25°C | Pseudomonas sp. | |
0.055 | - |
4-chlorobenzoyl-coenzyme A | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.069 | - |
4-hydroxybenzoyl-dithio-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.072 | - |
4-methylbenzoyl-CoA | mutant R257L, pH 7.5, 25°C | Pseudomonas sp. | |
0.09 | - |
CoA | mutant R257K, pH 7.5, 25°C | Pseudomonas sp. | |
0.14 | - |
CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
0.25 | - |
CoA | mutant R24K, pH 7.5, 25°C | Pseudomonas sp. | |
0.52 | - |
CoA | mutant R67K, pH 7.5, 25°C | Pseudomonas sp. | |
0.66 | - |
CoA | mutant R24L, pH 7.5, 25°C | Pseudomonas sp. | |
0.77 | - |
3'-dephospho-CoA | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
2.4 | - |
CoA | mutant R257L, pH 7.5, 25°C | Pseudomonas sp. | |
4.2 | - |
5'-ADP | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. | |
21 | - |
4-Chlorobenzoate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas sp. |