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Literature summary for 3.8.1.9 extracted from

  • Feng, Y.; Wang, Y.; Chu, H.; Fan, Y.; Cao, X.; Liu, Y.; Li, G.; Xue, S.
    Stereoselective catalysis controlled by a native leucine or variant isoleucine wing-gatekeeper in 2-haloacid dehalogenase (2019), FEBS Lett., 593, 308-318 .
    View publication on PubMed
Search for more literature for 3.8.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 cells Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals are obtained by the hanging-drop vapor-diffusion method. Crystals of the D205N mutant with D-2-chloropropionate are prepared by co-crystallization using the sitting drop method Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
D205N catalytically inactive variant Pseudomonas putida
L288A the mutant enzyme is active toward L-enantiomers. The catalytic efficiency is less than 30% of mutant enzyme L288I Pseudomonas putida
L288I the mutation enlarges the size of the channel and allows the enzyme to accommodate L-enantiomers (L-2-bromopropionate). The wing flip of I288 induces hydrophobic interactions with the L-enantiomer and directly affects the catalytic efficiency Pseudomonas putida
L288V kcat/Km of the L288V mutant displays over 80% of that of the L288I mutant Pseudomonas putida
M284F mutant enzyme shows no enantioselective changes in contrast to the wild-type enzyme Pseudomonas putida
N203A mutant enzyme shows no enantioselective changes in contrast to the wild-type enzyme Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.62
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
1.93
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
4.13
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
4.45
-
 D-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
4.88
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
6.03
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
8.07
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
32.8
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida Q52086
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-2-bromopropionate + H2O mutant enzyme L288I exhibits catalytic activity, no activity with wild-type enzyme Pseudomonas putida L-2-hydroxypropionate + bromide
-
 ?
D-2-chloropropionate + H2O mutant enzyme L288I exhibits catalytic activity, no activity with wild-type enzyme Pseudomonas putida L-2-hydroxypropionate + chloride
-
 ?
L-2-bromopropionate + H2O mutant enzyme L288I exhibits catalytic activity, no activity with wild-type enzyme Pseudomonas putida ?
-
 ?
L-2-chloropropionate + H2O mutant enzyme L288I exhibits catalytic activity, no activity with wild-type enzyme Pseudomonas putida ?
-
 ?

Synonyms

Synonyms Comment Organism
HadD AJ1
-
 Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.052
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
0.19
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
4.05
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
6.83
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
12.25
-
 D-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
14.83
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
34.17
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
253
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.03
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
0.1
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
0.12
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
0.123
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
1.65
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
1.84
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
5.18
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
5.6
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida