Information on EC 3.8.1.10 - 2-haloacid dehalogenase (configuration-inverting)

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.8.1.10
-
RECOMMENDED NAME
GeneOntology No.
2-haloacid dehalogenase (configuration-inverting)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-2-haloacid + H2O = (S)-2-hydroxyacid + halide
show the reaction diagram
(S)-2-haloacid + H2O = (R)-2-hydroxyacid + halide
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dehalogenation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
(S)-2-haloacid dehalogenase (configuration-inverting)
Dehalogenates both (S)- and (R)-2-haloalkanoic acids to the corresponding (R)- and (S)-hydroxyalkanoic acids, respectively, with inversion of configuration at C-2. The enzyme from Pseudomonas sp. 113 acts on 2-haloalkanoic acids whose carbon chain lengths are five or less. [See also EC 3.8.1.2 (S)-2-haloacid dehalogenase, EC 3.8.1.9 (R)-2-haloacid dehalogenase and EC 3.8.1.11 2-haloacid dehalogenase (configuration-retaining)]
CAS REGISTRY NUMBER
COMMENTARY hide
89511-96-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain KY
-
-
Manually annotated by BRENDA team
strain KY
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-
Manually annotated by BRENDA team
strain AJ1
-
-
Manually annotated by BRENDA team
strain YL
-
-
Manually annotated by BRENDA team
pv. tomato DC3000
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-2-chloropropionate + H2O
(S)-2-hydroxypropionate + chloride
show the reaction diagram
-
-
-
?
(R)-2-chloropropionic acid + H2O
(S)-2-hydroxypropionic acid + chloride
show the reaction diagram
(R)-2-haloacid + H2O
(S)-2-hydroxyacid + halide
show the reaction diagram
(S)-2-chloropropionate + H2O
(R)-2-hydroxypropionate + chloride
show the reaction diagram
-
-
-
?
(S)-2-chloropropionic acid + H2O
(R)-2-hydroxypropionic acid + chloride
show the reaction diagram
(S)-2-haloacid + H2O
(R)-2-hydroxyacid + halide
show the reaction diagram
2,2-dichloropropionate + H2O
?
show the reaction diagram
2,3-dichloropropionate + H2O
?
show the reaction diagram
2-bromo-2-methylpropionate + H2O
2-hydroxy-2-methylpropionate + bromide
show the reaction diagram
slow substrate for the wild-type enzyme, but inhibitory for enzyme mutant D194N
-
-
?
2-chloro-3-hydroxypropionate + H2O
glycerate + HCl
show the reaction diagram
2-chloro-n-butyrate + H2O
2-hydroxybutyrate + HCl
show the reaction diagram
D-2-chloropropionate + H2O
L-lactate + HCl
show the reaction diagram
D-2-monochloropropionate + H2O
L-lactate + HCl
show the reaction diagram
dichloroacetate + H2O
?
show the reaction diagram
DL-2-chloropropionate + H2O
DL-lactate + HCl
show the reaction diagram
L-2-chloropropionate + H2O
D-lactate + HCl
show the reaction diagram
L-2-monochloropropionate + H2O
D-lactate + HCl
show the reaction diagram
monobromoacetate + H2O
glycolate + HBr
show the reaction diagram
monochloroacetate + H2O
glycolate + HCl
show the reaction diagram
monoiodoacetate + H2O
glycolate + HI
show the reaction diagram
tribromoacetic acid + H2O
carbon monoxide + carbon dioxide + HBr
show the reaction diagram
-
-
-
?
trichloroacetate + H2O
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(R)-2-chloropropionic acid + H2O
(S)-2-hydroxypropionic acid + chloride
show the reaction diagram
(R)-2-haloacid + H2O
(S)-2-hydroxyacid + halide
show the reaction diagram
(S)-2-chloropropionic acid + H2O
(R)-2-hydroxypropionic acid + chloride
show the reaction diagram
(S)-2-haloacid + H2O
(R)-2-hydroxyacid + halide
show the reaction diagram
L-2-chloropropionate + H2O
D-lactate + HCl
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
-
complete inactivation
2-bromo-2-methylpropionate
an inhibitor of the D194N mutant enzyme and slow substrate of the wild-type enzyme
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D-2-chloropropionate
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inhibits dehalogenation of L-2-chloropropionate
diethyldicarbonate
-
significant inactivation
L-2-chloropropionate
-
inhibits dehalogenation of D-2-chloropropionate
Mg2+
the enzyme activity is suppressed by Mg2+
Mn2+
-
MnSO4
N-bromosuccinimide
-
significant inactivation
N-ethylmaleimide
-
-
p-chloromercuribenzoate
-
-
thiol reagents
-
not
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.2 - 4.8
D-2-chloropropionate
3.2
DL-2-chloropropionate
0.9 - 1.1
L-2-chloropropionate
4.8 - 5
Monochloroacetate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.2
D-2-chloropropionate
-
pH 9.5, 30°C
0.28
HgCl2
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pH 9.5, 30°C
6.5
L-2-chloropropionate
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pH 9.5, 30°C
0.51
ZnSO4
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pH 9.5, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.01
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D-2-chloropropinate, comparison with mutant enzyme
1.07
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L-2-chloropropinate, comparison with mutant enzyme
additional information
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-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
assay at room temperature
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.9
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isoelectric focusing
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
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2 * 28000, SDS-PAGE
34242
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2 * 34242, calculated from amino acid sequence
35000
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2 * 35000, SDS-PAGE
36000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant native and selenomethionine-labeled wild-type enzyme and recombinant D194N enzyme mutant complexed with inhibitor 2-bromo-2-methylpropionate, 15 mg/mL protein in 10 mM Hepes-NaOH, pH 7.5, is mixed with 3 M sodium formate, 5% glycerol v/v, as the reservoir solution, X-ray diffraction structure determination and analysis at 1.7-2.7 A resolution
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
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 10
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-
648274
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
15 min 50% loss of activity for D-2-chloropropionic acid and 40% loss of activity for L-2-chloropropionic acid
60
-
30 min 80% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 25 mM potassium phosphate buffer containing 50% glycerol, pH 7.5, 1 year
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
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recombinant enzyme DL-DEX Mb from Escherichia coli
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli, no significant sequence similarity between enzyme and L-2-haloacid dehalogenase is found, however, enzyme has significant sequence similarity with D-2-haloacid dehalogenase fom Pseudomonas putide AJ1
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gene located on pFL40
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recombinant expression of wild-type and mutant enzymes
recombinant expresssion of enzyme DL-DEX Mb in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D194N
site-directed mutagenesis
D180A
-
the mutation destabilizes the rotation of the nucleophile D10, fixes catalytic water around D10, and prevents K151 from approaching D10
D181A
-
the activity of mutant is similar to wild-type enzyme
D181E
-
the activity of mutant is similar to wild-type enzyme
D181R
-
the activity of mutant is similar to wild-type enzyme
K151A
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the mutant is almost inactive, the mutation destabilizes substrate orientation and affects the balance of the charge around the active site
K151R
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the mutant is almost inactive
S118A
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the mutant retains 30% of the activity of the wild type enzyme
D181A
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the activity of mutant is similar to wild-type enzyme
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D181E
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the activity of mutant is similar to wild-type enzyme
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D181R
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the activity of mutant is similar to wild-type enzyme
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D180A
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the mutation destabilizes the rotation of the nucleophile D10, fixes catalytic water around D10, and prevents K151 from approaching D10
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K151A
-
the mutant is almost inactive, the mutation destabilizes substrate orientation and affects the balance of the charge around the active site
-
K151R
-
the mutant is almost inactive
-
S118A
-
the mutant retains 30% of the activity of the wild type enzyme
-
D10A
site-directed mutagenesis, the D10A mutant shows over 50% reduced enzyme activity compared to the wild-type enzyme
D11A
site-directed mutagenesis, the D11A mutant shows highly reduced enzyme activity compared to the wild-type enzyme
D180A
site-directed mutagenesis, the D180A mutant shows over 50% reduced enzyme activity compared to the wild-type enzyme
D184A
site-directed mutagenesis, the mutant cannot be expressed recombinantly
D185A
site-directed mutagenesis, the mutant cannot be expressed recombinantly
D8A
site-directed mutagenesis, the D8A mutant retains about half the enzyme activity of the wild-type
K155A
site-directed mutagenesis, the mutant cannot be expressed recombinantly
T127A
site-directed mutagenesis, the mutant cannot be expressed recombinantly
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
the enzyme is of interest for their potential use in bioremediation and in the synthesis of industrial chemicals