Literature summary for 3.8.1.10 extracted from

Hou, Z.; Zhang, H.; Li, M.; Chang, W.
Structure of 2-haloacid dehalogenase from Pseudomonas syringae pv. tomato DC3000 (2013), Acta Crystallogr. Sect. D, 69, 1108-1114.

Application

Application	Comment	Organism
synthesis	the enzyme is of interest for their potential use in bioremediation and in the synthesis of industrial chemicals	Pseudomonas syringae

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of wild-type and mutant enzymes	Pseudomonas syringae

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme, sitting drop vapour diffusion method, mixing of equal volumes of 9 mg/ml protein in 50 mM Tris, pH 8.0, and 25 mM NaCl, and of reservoir solution consisting of 0.2 M MgCl2 hexahydrate, 20-26% PEG 4000, 0.1 M Tris, pH 8.5, and equilibration against 0.1 ml of reservoir solution, 4°C, overnight, soaking of crystals in KI, X-ray diffraction structure determination and analysis at 1.98-2.0 A resolution, single-wavelength anomalous dispersion method, modeling	Pseudomonas syringae

Crystallization (Comment)

Organism

purified enzyme, sitting drop vapour diffusion method, mixing of equal volumes of 9 mg/ml protein in 50 mM Tris, pH 8.0, and 25 mM NaCl, and of reservoir solution consisting of 0.2 M MgCl2 hexahydrate, 20-26% PEG 4000, 0.1 M Tris, pH 8.5, and equilibration against 0.1 ml of reservoir solution, 4°C, overnight, soaking of crystals in KI, X-ray diffraction structure determination and analysis at 1.98-2.0 A resolution, single-wavelength anomalous dispersion method, modeling

Pseudomonas syringae

Protein Variants

Protein Variants	Comment	Organism
D10A	site-directed mutagenesis, the D10A mutant shows over 50% reduced enzyme activity compared to the wild-type enzyme	Pseudomonas syringae
D11A	site-directed mutagenesis, the D11A mutant shows highly reduced enzyme activity compared to the wild-type enzyme	Pseudomonas syringae
D180A	site-directed mutagenesis, the D180A mutant shows over 50% reduced enzyme activity compared to the wild-type enzyme	Pseudomonas syringae
D184A	site-directed mutagenesis, the mutant cannot be expressed recombinantly	Pseudomonas syringae
D185A	site-directed mutagenesis, the mutant cannot be expressed recombinantly	Pseudomonas syringae
D8A	site-directed mutagenesis, the D8A mutant retains about half the enzyme activity of the wild-type	Pseudomonas syringae
K155A	site-directed mutagenesis, the mutant cannot be expressed recombinantly	Pseudomonas syringae
T127A	site-directed mutagenesis, the mutant cannot be expressed recombinantly	Pseudomonas syringae

Inhibitors

Inhibitors	Comment	Organism	Structure
Mg2+	the enzyme activity is suppressed by Mg2+	Pseudomonas syringae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
(R)-2-haloacid + H2O	Pseudomonas syringae	-	(S)-2-hydroxyacid + halide	-	?
(S)-2-haloacid + H2O	Pseudomonas syringae	-	(R)-2-hydroxyacid + halide	-	?
additional information	Pseudomonas syringae	2-haloacid dehalogenases catalyse the hydrolytic dehalogenation of 2-haloalkanoic acids, cleaving the carbon-halide bond at the Calpha-atom position and releasing a halogen atom	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas syringae	Q88B12	pv. tomato DC3000	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(R)-2-haloacid + H2O	-	Pseudomonas syringae	(S)-2-hydroxyacid + halide	-	?
(S)-2-haloacid + H2O	-	Pseudomonas syringae	(R)-2-hydroxyacid + halide	-	?
additional information	2-haloacid dehalogenases catalyse the hydrolytic dehalogenation of 2-haloalkanoic acids, cleaving the carbon-halide bond at the Calpha-atom position and releasing a halogen atom	Pseudomonas syringae	?	-	?
additional information	enzyme ps-2-HAD reveals its capacity to catalyse the dehalogenation of both L- and D-substrates	Pseudomonas syringae	?	-	?

Synonyms

Synonyms	Comment	Organism
2-HAD	-	Pseudomonas syringae
ps-2-HAD	-	Pseudomonas syringae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Pseudomonas syringae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.2	-	assay at	Pseudomonas syringae

General Information

General Information	Comment	Organism
evolution	Based on their substrate specificities and the configurations of their products, 2-HADs are classified into three types. DL-2-HADs catalyse the dehalogenation of both D- and L-2-haloalkanoic acids, inverting the stereochemistry at the Calpha-atom position and producing the corresponding L- and D-products, respectively. D- and L-2-HADs act specifically on only one enantiomer and cause inversion of Calpha-atom stereochemistry in the hydroxyalkanoic acid product. Enzymatic activity analysis of Pseudomonas syringae 2-HAD reveals its capacity to catalyse the dehalogenation of both L- and D-substrates, but the structure of ps-2-HAD is completely different from that of DehI, which is the only DL-2-HAD enzyme that is structurally characterized, bps-2-HAD shows similar overall folding to L-HADs. Thus ps-2-HAD has a distinct active site and a unique catalytic behaviour compared with other HADs	Pseudomonas syringae
additional information	either the magnesium-binding cavity at least partially overlaps with the active site of ps-2-HAD or the coordinated residues or water molecules are involved directly in the catalytic reaction	Pseudomonas syringae