Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, recombinant expression | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
D130A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
D184A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
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. |
D45A | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
H128Q | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
H157Q | site-directed mutagenesis, inactive mutant | Pseudomonas sp. |
H63Q | site-directed mutagenesis, the Km for the mutant Chd increases to 0.154 mM compared to the wild-type | Pseudomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | complete inhibition, recovered by the subsequent supplementation with Zn2+ | Pseudomonas sp. | |
diethyl dicarbonate | complete inhibition | Pseudomonas sp. | |
N-bromosuccinimide | complete inhibition | Pseudomonas sp. | |
SDS | 90% inhibition | Pseudomonas sp. | |
Triton X-100 | 50% inhibition | Pseudomonas sp. | |
Tween 80 | 60% inhibition | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.112 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | no activating effects by Cu2+, Ba2+, Mg2+, or Ni2+ | Pseudomonas sp. | |
Zn2+ | activates | Pseudomonas sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33886 | - |
1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry | Pseudomonas sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4,5,6-tetrachloroisophthalonitrile + H2O | Pseudomonas sp. | - |
4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
2,4,5,6-tetrachloroisophthalonitrile + H2O | Pseudomonas sp. CTN-3 | - |
4-hydroxy-trichloroisophthalonitrile + chloride | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | C7EW69 | a 2,4,5,6-tetrachloroisophthalonitrile (chlorothalonil)-degrading strain | - |
Pseudomonas sp. CTN-3 | C7EW69 | a 2,4,5,6-tetrachloroisophthalonitrile (chlorothalonil)-degrading strain | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Pseudomonas sp. |
Storage Stability | Organism |
---|---|
-20°C, purified enzyme,50 mM PBS, pH 7.0, 10% glycerol, no loss of Chd activity within 3 months | Pseudomonas sp. |
4°C, purified enzyme, 3-5% of Chd activity is lost within 8 weeks | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4,5,6-tetrachloroisophthalonitrile + H2O | - |
Pseudomonas sp. | 4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
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 | ? | |
2,4,5,6-tetrachloroisophthalonitrile + H2O | - |
Pseudomonas sp. CTN-3 | 4-hydroxy-trichloroisophthalonitrile + chloride | - |
? | |
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 | ? | |
additional information | no activity with chlorobenzene, 4-dichlorobenzene, 4-chloronitrobenzene, pentachloronitrobenzene, 4-chlorobenzoate, 4-chlorophenylacetic acid, 3-chloroaniline, 4-chloroaniline, and pentachlorophenol | Pseudomonas sp. | ? | - |
? | |
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 | Comment | Organism |
---|---|---|
monomer | 1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry | Pseudomonas sp. |
Synonyms | Comment | Organism |
---|---|---|
4-chlorobenzoyl-CoA dehalogenase | - |
Pseudomonas sp. |
4-chlorobenzoyl-coenzyme A dehalogenase | - |
Pseudomonas sp. |
CHD | - |
Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Pseudomonas sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
Chd is fairly stable up to | Pseudomonas sp. |
50 | - |
purified enzyme, 95% activity remaining after 10 min, 18% after 1 h | Pseudomonas sp. |
60 | - |
purified Chd, complete loss of activity after 1 h | Pseudomonas sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
207 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Pseudomonas sp. |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | purified Chd, 95% activity remaining after 30 min | Pseudomonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
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. |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudomonas sp. | sequence calculation | - |
4.13 |
General Information | Comment | Organism |
---|---|---|
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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.185 | - |
2,4,5,6-tetrachloroisophthalonitrile | pH 7.0, 50°C | Pseudomonas sp. |