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Literature summary extracted from
- A novel hydrolytic dehalogenase for the chlorinated aromatic compound chlorothalonil (2010), J. Bacteriol., 192, 2737-2745.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.8.1.7 | DNA and amino acid sequence determination and analysis, recombinant expression | Pseudomonas sp. |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.8.1.7 | D130A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
| 3.8.1.7 | D184A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
| 3.8.1.7 | D337A | site-directed mutagenesis, the Km for the mutant Chd increases to 0.176 mM, 50% transformation activity compared to the wild-type Chd | Pseudomonas sp. |
| 3.8.1.7 | D45A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
| 3.8.1.7 | H128Q | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
| 3.8.1.7 | H157Q | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
| 3.8.1.7 | H63Q | site-directed mutagenesis, the Km for the mutant Chd increases to 0.154 mM compared to the wild-type | Pseudomonas sp. |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.8.1.7 | 1,10-phenanthroline | complete inhibition, recovered by the subsequent supplementation with Zn2+ | Pseudomonas sp. |  |
| 3.8.1.7 | diethyl dicarbonate | complete inhibition | Pseudomonas sp. | |
| 3.8.1.7 | N-bromosuccinimide | complete inhibition | Pseudomonas sp. | |
| 3.8.1.7 | SDS | 90% inhibition | Pseudomonas sp. | |
| 3.8.1.7 | Triton X-100 | 50% inhibition | Pseudomonas sp. | |
| 3.8.1.7 | Tween 80 | 60% inhibition | Pseudomonas sp. | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.8.1.7 | 0.112 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.8.1.7 | additional information | no activating effects by Cu2+, Ba2+, Mg2+, or Ni2+ | Pseudomonas sp. | |
| 3.8.1.7 | Zn2+ | activates | Pseudomonas sp. | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.8.1.7 | 33886 | - |
1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry | Pseudomonas sp. |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | Pseudomonas sp. | - |
4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | Pseudomonas sp. CTN-3 | - |
4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.8.1.7 | Pseudomonas sp. | C7EW69 | a 2,4,5,6-tetrachloroisophthalonitrile (chlorothalonil)-degrading strain | - |
| 3.8.1.7 | Pseudomonas sp. CTN-3 | C7EW69 | a 2,4,5,6-tetrachloroisophthalonitrile (chlorothalonil)-degrading strain | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.8.1.7 | recombinant enzyme | Pseudomonas sp. |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 3.8.1.7 | -20°C, purified enzyme,50 mM PBS, pH 7.0, 10% glycerol, no loss of Chd activity within 3 months | Pseudomonas sp. |
| 3.8.1.7 | 4°C, purified enzyme, 3-5% of Chd activity is lost within 8 weeks | Pseudomonas sp. |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | - |
Pseudomonas sp. | 4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | a regiospecific hydroxyl substitution at the 4-chlorine atom of chlorothalonil. Histidines 128 and 157, serine 126, aspartates 45, 130 and 184, and tryptophan 241 are essential for the dehalogenase activity | Pseudomonas sp. | 4-hydroxy-trichloroisophthalonitrile + chloride | product identified by reverse-phase HPLC, tandem mass spectrometry, and NMR | ? | |
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | - |
Pseudomonas sp. CTN-3 | 4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
| 3.8.1.7 | 2,4,5,6-tetrachloroisophthalonitrile + H2O | a regiospecific hydroxyl substitution at the 4-chlorine atom of chlorothalonil. Histidines 128 and 157, serine 126, aspartates 45, 130 and 184, and tryptophan 241 are essential for the dehalogenase activity | Pseudomonas sp. CTN-3 | 4-hydroxy-trichloroisophthalonitrile + chloride | product identified by reverse-phase HPLC, tandem mass spectrometry, and NMR | ? | |
| 3.8.1.7 | additional information | no activity with chlorobenzene, 4-dichlorobenzene, 4-chloronitrobenzene, pentachloronitrobenzene, 4-chlorobenzoate, 4-chlorophenylacetic acid, 3-chloroaniline, 4-chloroaniline, and pentachlorophenol | Pseudomonas sp. | ? | - |
? | |
| 3.8.1.7 | additional information | no activity with chlorobenzene, 4-dichlorobenzene, 4-chloronitrobenzene, pentachloronitrobenzene, 4-chlorobenzoate, 4-chlorophenylacetic acid, 3-chloroaniline, 4-chloroaniline, and pentachlorophenol | Pseudomonas sp. CTN-3 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.8.1.7 | monomer | 1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry | Pseudomonas sp. |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.8.1.7 | 4-chlorobenzoyl-CoA dehalogenase | - |
Pseudomonas sp. |
| 3.8.1.7 | 4-chlorobenzoyl-coenzyme A dehalogenase | - |
Pseudomonas sp. |
| 3.8.1.7 | CHD | - |
Pseudomonas sp. |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.8.1.7 | 50 | - |
- |
Pseudomonas sp. |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.8.1.7 | 40 | - |
Chd is fairly stable up to | Pseudomonas sp. |
| 3.8.1.7 | 50 | - |
purified enzyme, 95% activity remaining after 10 min, 18% after 1 h | Pseudomonas sp. |
| 3.8.1.7 | 60 | - |
purified Chd, complete loss of activity after 1 h | Pseudomonas sp. |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.8.1.7 | 207 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.8.1.7 | 7 | - |
- |
Pseudomonas sp. |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.8.1.7 | 6 | 9 | purified Chd, 95% activity remaining after 30 min | Pseudomonas sp. |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.8.1.7 | additional information | Chd dehalogenates chlorothalonil under anaerobic and aerobic conditions and does not require the presence of cofactors such as CoA and ATP | Pseudomonas sp. |
pI Value
| EC Number | Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|---|
| 3.8.1.7 | Pseudomonas sp. | sequence calculation | - |
4.13 |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.8.1.7 | evolution | Chd contains a putative conserved domain of the metallo-beta-lactamase superfamily and shows the highest identity with several metallohydrolases | Pseudomonas sp. |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.8.1.7 | 0.185 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. | |
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