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Literature summary for 3.8.1.2 extracted from

  • Novak, H.R.; Sayer, C.; Isupov, M.N.; Paszkiewicz, K.; Gotz, D.; Spragg, A.M.; Littlechild, J.A.
    Marine Rhodobacteraceae L-haloacid dehalogenase contains a novel His/Glu dyad that could activate the catalytic water (2013), FEBS J., 280, 1664-1680.
    View publication on PubMed

Cloned(Commentary)

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 [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 [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/ Products (Substrates)

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 Stability

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

Organism UniProt Comment Textmining
Rhodobacteraceae bacterium UDC319 M9P6K0 gene DehRhb
-

Purification (Commentary)

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

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 and Products (Substrate)

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

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Rhodobacteraceae bacterium UDC319

Temperature Stability [°C]

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

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