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Literature summary 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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.8.1.9 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

EC Number Protein Variants Comment Organism
3.8.1.9 D205N catalytically inactive variant Pseudomonas putida
3.8.1.9 L288A the mutant enzyme is active toward L-enantiomers. The catalytic efficiency is less than 30% of mutant enzyme L288I Pseudomonas putida
3.8.1.9 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
3.8.1.9 L288V kcat/Km of the L288V mutant displays over 80% of that of the L288I mutant Pseudomonas putida
3.8.1.9 M284F mutant enzyme shows no enantioselective changes in contrast to the wild-type enzyme Pseudomonas putida
3.8.1.9 N203A mutant enzyme shows no enantioselective changes in contrast to the wild-type enzyme Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.8.1.9 1.62
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 1.93
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
3.8.1.9 4.13
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 4.45
-
 D-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 4.88
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
3.8.1.9 6.03
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 8.07
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 32.8
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
3.8.1.9 Pseudomonas putida Q52086
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.8.1.9
-
 Pseudomonas putida

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
3.8.1.9 HadD AJ1
-
 Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.8.1.9 0.052
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 0.19
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
3.8.1.9 4.05
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 6.83
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 12.25
-
 D-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 14.83
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 34.17
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 253
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.8.1.9 0.03
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 0.1
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
3.8.1.9 0.12
-
 D-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 0.123
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 1.65
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288A Pseudomonas putida
3.8.1.9 1.84
-
 L-2-chloropropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida
3.8.1.9 5.18
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288V Pseudomonas putida
3.8.1.9 5.6
-
 L-2-bromopropionate pH 10.0, 30°C, mutant enzyme L288I Pseudomonas putida