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Literature summary extracted from

  • Hasan, K.; Gora, A.; Brezovsky, J.; Chaloupkova, R.; Moskalikova, H.; Fortova, A.; Nagata, Y.; Damborsky, J.; Prokop, Z.
    The effect of a unique halide-stabilizing residue on the catalytic properties of haloalkane dehalogenase DatA from Agrobacterium tumefaciens C58 (2013), FEBS J., 280, 3149-3159.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
3.8.1.5 Y109W site-directed mutagenesis, structure and catalytic properties by spectroscopy and pre-steady-state kinetic experiments, quantum mechanical and molecular dynamics calculations. The mutation affects the substrate specificity of the enzyme and reduced its KM for selected halogenated substrates by a factor of 2-4, without impacting the rate-determining hydrolytic step Agrobacterium tumefaciens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.8.1.5 additional information
-
 additional information pre-steady-state and steady-state kinetics, both the wild-type and the mutant yield hyperbolic kinetic profiles for their reactions with 1,3-dibromopropane and 1-bromohexane, suggesting classical Michaelis-Menten kinetics. The sigmoidal profiles obtained in experiments using 1,3-diiodopropane, 1-chlorohexane and 1-bromobutane indicate that these substrates are hydrolyzed via a cooperative Hill mechanism, KM values for the mutant are lower than for the wild-type enzyme. Substrate binding and the cleavage of the carbon-halogen bond are fast steps, and that hydrolysis of the alkyl-enzyme intermediate is the rate-determining step in the conversion of 1-bromobutane by both enzymes Agrobacterium tumefaciens
3.8.1.5 0.03
-
 1-bromohexane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.06
-
 1-Chlorohexane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.07
-
 1-Chlorohexane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.14
-
 1-bromohexane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.16
-
 1,3-diiodopropane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.18
-
 1-bromobutane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.2
-
 1,3-diiodopropane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.53
-
 1-bromobutane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.82
-
 1,3-dibromopropane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 2.2
-
 1,3-dibromopropane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 4
-
 1,3-Dichloropropane above, recombinant wild-type enzyme and mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens

Organism

EC Number Organism UniProt Comment Textmining
3.8.1.5 Agrobacterium tumefaciens E2RV69 gene datA
-
3.8.1.5 Agrobacterium tumefaciens C58 / ATCC 33970 E2RV69 gene datA
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.8.1.5 1,2,3-tribromopropane + H2O high activity Agrobacterium tumefaciens 2,3-dibromopropanol + bromide
-
 ?
3.8.1.5 1,2-dibromo-3-chloropropane + H2O high activity Agrobacterium tumefaciens 2-bromo-3-chloropropanol + bromide
-
 ?
3.8.1.5 1,2-dibromoethane + H2O high activity Agrobacterium tumefaciens 2-bromoethanol + bromide
-
 ?
3.8.1.5 1,2-dibromoethane + H2O high activity Agrobacterium tumefaciens C58 / ATCC 33970 2-bromoethanol + bromide
-
 ?
3.8.1.5 1,2-dibromopropane + H2O high activity Agrobacterium tumefaciens 2-bromopropanol + bromide
-
 ?
3.8.1.5 1,3-dibromopropane + H2O
-
 Agrobacterium tumefaciens 3-bromopropanol + bromide
-
 ?
3.8.1.5 1,3-dichloropropane + H2O
-
 Agrobacterium tumefaciens 3-chloropropanol + chloride
-
 ?
3.8.1.5 1,3-diiodopropane + H2O
-
 Agrobacterium tumefaciens 3-iodopropanol + iodide
-
 ?
3.8.1.5 1,5-dichloropentane + H2O
-
 Agrobacterium tumefaciens 5-chloropentanol + chloride
-
 ?
3.8.1.5 1-bromo-2-chloroethane + H2O high activity Agrobacterium tumefaciens 2-chloroethanol + bromide
-
 ?
3.8.1.5 1-bromobutane + H2O
-
 Agrobacterium tumefaciens butanol + bromide
-
 ?
3.8.1.5 1-bromobutane + H2O
-
 Agrobacterium tumefaciens C58 / ATCC 33970 butanol + bromide
-
 ?
3.8.1.5 1-bromohexane + H2O
-
 Agrobacterium tumefaciens 1-hexanol + bromide
-
 ?
3.8.1.5 1-chlorohexane + H2O
-
 Agrobacterium tumefaciens 1-hexanol + chloride
-
 ?
3.8.1.5 1-iodobutane + H2O
-
 Agrobacterium tumefaciens butanol + iodide
-
 ?
3.8.1.5 1-iodobutane + H2O
-
 Agrobacterium tumefaciens C58 / ATCC 33970 butanol + iodide
-
 ?
3.8.1.5 1-iodopropane + H2O
-
 Agrobacterium tumefaciens propanol + iodide
-
 ?
3.8.1.5 1-iodopropane + H2O
-
 Agrobacterium tumefaciens C58 / ATCC 33970 propanol + iodide
-
 ?
3.8.1.5 2-iodobutane + H2O
-
 Agrobacterium tumefaciens 2-butanol + iodide
-
 ?
3.8.1.5 3-chloro-2-methylpropene + H2O high activity Agrobacterium tumefaciens ?
-
 ?
3.8.1.5 4-bromobutyronitrile + H2O high activity Agrobacterium tumefaciens ?
-
 ?
3.8.1.5 bis(2-chloroethyl)ether + H2O high activity Agrobacterium tumefaciens ?
-
 ?
3.8.1.5 additional information substrate specificity, substrate docking and molecular modelling, overview. No activity with 2,3-dichloropropene, chlorocyclohexane, 1,2,3-trichloropropane, 1,2-dichloropropane, and 1,2-dichloroethane Agrobacterium tumefaciens ?
-
 ?
3.8.1.5 additional information substrate specificity, substrate docking and molecular modelling, overview. No activity with 2,3-dichloropropene, chlorocyclohexane, 1,2,3-trichloropropane, 1,2-dichloropropane, and 1,2-dichloroethane Agrobacterium tumefaciens C58 / ATCC 33970 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.8.1.5 DatA
-
 Agrobacterium tumefaciens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.8.1.5 37
-
 assay at Agrobacterium tumefaciens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.8.1.5 0.01
-
 1,3-Dichloropropane below, recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.012
-
 1,3-Dichloropropane below, recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.04
-
 1-Chlorohexane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.07
-
 1-Chlorohexane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.46
-
 1-bromohexane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.71
-
 1-bromobutane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 0.72
-
 1,3-diiodopropane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 1.2
-
 1,3-diiodopropane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 1.3
-
 1,3-dibromopropane recombinant mutant Y109W, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 1.5
-
 1,3-dibromopropane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 1.5
-
 1-bromobutane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens
3.8.1.5 1.7
-
 1-bromohexane recombinant wild-type enzyme, pH 8.6, 37°C Agrobacterium tumefaciens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.8.1.5 8.6
-
 assay at Agrobacterium tumefaciens

General Information

EC Number General Information Comment Organism
3.8.1.5 evolution the enzyme possesses a unique halide-stabilizing tyrosine residue, Y109, in place of the conventional tryptophan. Enzyme DatA is unique in stabilizing its substrate in the active site using only a single hydrogen bond, which is a distinct paradigm in catalysis by this enzyme family Agrobacterium tumefaciens