Protein Variants | Comment | Organism |
---|---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | |
0.03 | - |
1-bromohexane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.06 | - |
1-Chlorohexane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.07 | - |
1-Chlorohexane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.14 | - |
1-bromohexane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.16 | - |
1,3-diiodopropane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.18 | - |
1-bromobutane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.2 | - |
1,3-diiodopropane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.53 | - |
1-bromobutane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.82 | - |
1,3-dibromopropane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
2.2 | - |
1,3-dibromopropane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
4 | - |
1,3-Dichloropropane | above, recombinant wild-type enzyme and mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium tumefaciens | E2RV69 | gene datA | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | E2RV69 | gene datA | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,2,3-tribromopropane + H2O | high activity | Agrobacterium tumefaciens | 2,3-dibromopropanol + bromide | - |
? | |
1,2-dibromo-3-chloropropane + H2O | high activity | Agrobacterium tumefaciens | 2-bromo-3-chloropropanol + bromide | - |
? | |
1,2-dibromoethane + H2O | high activity | Agrobacterium tumefaciens | 2-bromoethanol + bromide | - |
? | |
1,2-dibromoethane + H2O | high activity | Agrobacterium tumefaciens C58 / ATCC 33970 | 2-bromoethanol + bromide | - |
? | |
1,2-dibromopropane + H2O | high activity | Agrobacterium tumefaciens | 2-bromopropanol + bromide | - |
? | |
1,3-dibromopropane + H2O | - |
Agrobacterium tumefaciens | 3-bromopropanol + bromide | - |
? | |
1,3-dichloropropane + H2O | - |
Agrobacterium tumefaciens | 3-chloropropanol + chloride | - |
? | |
1,3-diiodopropane + H2O | - |
Agrobacterium tumefaciens | 3-iodopropanol + iodide | - |
? | |
1,5-dichloropentane + H2O | - |
Agrobacterium tumefaciens | 5-chloropentanol + chloride | - |
? | |
1-bromo-2-chloroethane + H2O | high activity | Agrobacterium tumefaciens | 2-chloroethanol + bromide | - |
? | |
1-bromobutane + H2O | - |
Agrobacterium tumefaciens | butanol + bromide | - |
? | |
1-bromobutane + H2O | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | butanol + bromide | - |
? | |
1-bromohexane + H2O | - |
Agrobacterium tumefaciens | 1-hexanol + bromide | - |
? | |
1-chlorohexane + H2O | - |
Agrobacterium tumefaciens | 1-hexanol + chloride | - |
? | |
1-iodobutane + H2O | - |
Agrobacterium tumefaciens | butanol + iodide | - |
? | |
1-iodobutane + H2O | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | butanol + iodide | - |
? | |
1-iodopropane + H2O | - |
Agrobacterium tumefaciens | propanol + iodide | - |
? | |
1-iodopropane + H2O | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | propanol + iodide | - |
? | |
2-iodobutane + H2O | - |
Agrobacterium tumefaciens | 2-butanol + iodide | - |
? | |
3-chloro-2-methylpropene + H2O | high activity | Agrobacterium tumefaciens | ? | - |
? | |
4-bromobutyronitrile + H2O | high activity | Agrobacterium tumefaciens | ? | - |
? | |
bis(2-chloroethyl)ether + H2O | high activity | Agrobacterium tumefaciens | ? | - |
? | |
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 | ? | - |
? | |
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 | Comment | Organism |
---|---|---|
DatA | - |
Agrobacterium tumefaciens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Agrobacterium tumefaciens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
1,3-Dichloropropane | below, recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.012 | - |
1,3-Dichloropropane | below, recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.04 | - |
1-Chlorohexane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.07 | - |
1-Chlorohexane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.46 | - |
1-bromohexane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.71 | - |
1-bromobutane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
0.72 | - |
1,3-diiodopropane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
1.2 | - |
1,3-diiodopropane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
1.3 | - |
1,3-dibromopropane | recombinant mutant Y109W, pH 8.6, 37°C | Agrobacterium tumefaciens | |
1.5 | - |
1,3-dibromopropane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
1.5 | - |
1-bromobutane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens | |
1.7 | - |
1-bromohexane | recombinant wild-type enzyme, pH 8.6, 37°C | Agrobacterium tumefaciens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.6 | - |
assay at | Agrobacterium tumefaciens |
General Information | Comment | Organism |
---|---|---|
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 |