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Literature summary for 3.8.1.7 extracted from

  • Wang, G.; Li, R.; Li, S.; Jiang, J.
    A novel hydrolytic dehalogenase for the chlorinated aromatic compound chlorothalonil (2010), J. Bacteriol., 192, 2737-2745.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.8.1.7

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, recombinant expression Pseudomonas sp.

Protein Variants

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

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 [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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
33886
-
 1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry Pseudomonas sp.

Natural Substrates/ Products (Substrates)

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

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 (Commentary)

Purification (Comment) Organism
recombinant enzyme Pseudomonas sp.

Storage Stability

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 and Products (Substrate)

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

Subunits Comment Organism
monomer 1 x 36000, SDS-PAGE, 1 * 33886, mass spectrometry Pseudomonas sp.

Synonyms

Synonyms Comment Organism
4-chlorobenzoyl-CoA dehalogenase
-
 Pseudomonas sp.
4-chlorobenzoyl-coenzyme A dehalogenase
-
 Pseudomonas sp.
CHD
-
 Pseudomonas sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Pseudomonas sp.

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Pseudomonas sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 9 purified Chd, 95% activity remaining after 30 min Pseudomonas sp.

Cofactor

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.

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas sp. sequence calculation
-
 4.13

General Information

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 [mM/s]

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.