Cloned (Comment) | Organism |
---|---|
recombinant overexpression of wild-tye and mutant enzymes in Escherichia coli strain Rosetta (DE3) | Agrobacterium tumefaciens |
Crystallization (Comment) | Organism |
---|---|
purified wild-type and mutant Y109W enzymes, sitting drop vapor diffusion method, mixing of 10 mg/mL protein in 10 mM Tris-HCl, pH 8.0, and 1 mM DTT with an equal volume of reservoir solution containing 0.1 M CHES, pH 8.6, 1.0 M potassium sodium tartrate, 0.2 M lithium sulfate, 0.01 M barium chloride, 20°C X-ray diffraction structure determination and analysis at 1.70 A and 1.95 A resolution, respectively, molecular replacement method using the crystal structure of Rv2579 from Mycobacterium tuberculosis, PDB ID 2qvb | Agrobacterium tumefaciens |
Protein Variants | Comment | Organism |
---|---|---|
Y109W | site-directed mutagenesis, crystal structure determination | Agrobacterium tumefaciens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
N-cyclohexyl-2-aminoethanesulfonic acid | CHES, competitive inhibition, CHES binds to the active site of DatA, the sulfate moiety of CHES is bound by Asn43, and the other parts are bound by Asp108 and His274 of the catalytic triad | Agrobacterium tumefaciens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Agrobacterium tumefaciens | enzyme DatA hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton | ? | - |
? | |
additional information | Agrobacterium tumefaciens C58 / ATCC 33970 | enzyme DatA hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium tumefaciens | E2RV69 | - |
- |
Agrobacterium tumefaciens C58 / ATCC 33970 | E2RV69 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-tye and mutant enzymes from Escherichia coli strain Rosetta (DE3) | Agrobacterium tumefaciens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
1-haloalkane + H2O = a primary alcohol + halide | structural basis for its reaction mechanism | Agrobacterium tumefaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,3-dibromopropane + H2O | - |
Agrobacterium tumefaciens | 3-bromopropanol + bromide | - |
? | |
1,3-dibromopropane + H2O | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | 3-bromopropanol + bromide | - |
? | |
additional information | enzyme DatA hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton | Agrobacterium tumefaciens | ? | - |
? | |
additional information | enzyme DatA hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton | Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DatA | - |
Agrobacterium tumefaciens |
HLD | - |
Agrobacterium tumefaciens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Agrobacterium tumefaciens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
95 | - |
purified recombinant enzyme, 5 min, inactivation | Agrobacterium tumefaciens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Agrobacterium tumefaciens |
General Information | Comment | Organism |
---|---|---|
evolution | haloalkane dehalogenase DatA from Agrobacterium tumefaciens strain C58 belongs to the HLD-II subfamily. Enzyme DatA possesses a unique Asn-Tyr pair instead of the Asn-Trp pair conserved among the subfamily members, which keeps the released halide ion stable | Agrobacterium tumefaciens |
additional information | docking models of enzyme DatA complexed with 1,3-dibromopropane and 2-bromohexane, the docking pose shows the highest score for each enantiomer of 2-bromohexane, only the (R)-form of 2-bromohexane can be in a near-attack conformation for the SN2 reaction. Location of the active site, and overall structure of the wild-type DatA, overview | Agrobacterium tumefaciens |