Cloned (Comment) | Organism |
---|---|
gene DehRhb, DNA and amino acid sequence determination and analysis, recombinant overexpression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)-RIPL | Rhodobacteraceae bacterium UDC319 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.72 | - |
2-bromoacetic acid | recombinant enzyme, pH and temperature not specified in the publication | Rhodobacteraceae bacterium UDC319 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
25000 | - |
2 * 25000, SDS-PAGE, 2 * 25600, about, sequence calculation | Rhodobacteraceae bacterium UDC319 |
25600 | - |
2 * 25000, SDS-PAGE, 2 * 25600, about, sequence calculation | Rhodobacteraceae bacterium UDC319 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-2-haloacid + H2O | Rhodobacteraceae bacterium UDC319 | - |
(R)-2-hydroxyacid + halide | - |
? |
Organic Solvent | Comment | Organism |
---|---|---|
additional information | enzyme DehRhb has a relatively high tolerance to organic solvents, i.e. in ethanol, methanol, acetonitrile, or dimethylsulfoxide | Rhodobacteraceae bacterium UDC319 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodobacteraceae bacterium UDC319 | M9P6K0 | gene DehRhb | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3)-RIPL by nickel affinity chromatography and gel filtration | Rhodobacteraceae bacterium UDC319 |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-2-haloacid + H2O = (R)-2-hydroxyacid + halide | structure-function analysis, catalytic mechanism, modeling. Asp181 is proposed to participate in the activation of the catalytic water molecule required for hydrolysis of the ester intermediate, Asp186 and Lys157 in DehRhb are responsible for deprotonation of the catalytic water molecule in the second part of the enzyme reaction. In DehRhb, the catalytic water molecule is coordinated by His183 and the conserved Asp186. This residue makes an ion pair with the conserved Lys157. The His183 makes an H-bond with the catalytic Asp18 on one side and an ion pair with Glu21 on the other side. This forms a catalytic triad similar to that observed in the haloalkane dehalogenases | Rhodobacteraceae bacterium UDC319 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-2-bromopropionic acid + H2O | - |
Rhodobacteraceae bacterium UDC319 | (R)-2-hydroxypropionate + bromide | - |
? | |
(S)-2-chloropropionic acid + H2O | - |
Rhodobacteraceae bacterium UDC319 | (R)-2-hydroxypropionate + chloride | - |
? | |
(S)-2-haloacid + H2O | - |
Rhodobacteraceae bacterium UDC319 | (R)-2-hydroxyacid + halide | - |
? | |
2-bromoacetic acid + H2O | - |
Rhodobacteraceae bacterium UDC319 | 2-hydroxyacetate + bromide | - |
? | |
2-chloroacetic acid + H2O | - |
Rhodobacteraceae bacterium UDC319 | 2-hydroxyacetate + chloride | - |
? | |
dichloroacetic acid + H2O | low activity | Rhodobacteraceae bacterium UDC319 | ? | - |
? | |
additional information | no activity with 2-chlorobutyric acid, trichloroacetic acid, 3-bromopropionic acid, fluoroacetate, (R)-2-chloropropionic acid, (R)-2-bromoopropionic acid, 1,2-dichloropropane, and 1-bromoheptane | Rhodobacteraceae bacterium UDC319 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 25000, SDS-PAGE, 2 * 25600, about, sequence calculation | Rhodobacteraceae bacterium UDC319 |
More | enzyme structure comparisons, overview. The two subunits of DehRhb are related by a molecular twofold axis to which the a2 helices from each subunit are parallel, folding of the DehRhb monomer and active site structure, structure modeling | Rhodobacteraceae bacterium UDC319 |
Synonyms | Comment | Organism |
---|---|---|
L-HAD | - |
Rhodobacteraceae bacterium UDC319 |
L-haloacid dehalogenase | - |
Rhodobacteraceae bacterium UDC319 |
Rhodobacteraceae family L-haloacid dehalogenase | - |
Rhodobacteraceae bacterium UDC319 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Rhodobacteraceae bacterium UDC319 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 70 | recombinant enzyme DehRhb has a melting temperature of 67°C, it is completely stable at 30°C, loses 10% activity at 40° and 30% at 55°C after 90mmin, 85% activity are remaining after 90 min at 50°C, 14% activity remaining after 90 min at 60°C | Rhodobacteraceae bacterium UDC319 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the HAD enzyme superfamily, which use an aspartate group as the nucleophile in their catalytic mechanism.. The HAD superfamily of enzymes also includes phosphate monoesterases, ATPases, phosphonoacetaldehyde hydrolases and phosphomutases | Rhodobacteraceae bacterium UDC319 |
additional information | folding of the DehRhb monomer and active site structure with catalytic residue Asp18, overall structure modeling. The halogen cradle in enzyme DehRhb is formed by the side chains of Phe47, Ile51, Phe66, Asn125 and Trp185 | Rhodobacteraceae bacterium UDC319 |