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Literature summary for 4.1.3.43 extracted from

  • Baker, P.; Seah, S.Y.K.
    Rational design of stereoselectivity in the class II pyruvate aldolase BphI (2012), J. Am. Chem. Soc., 134, 507-513.
    View publication on PubMed
Search for more literature for 4.1.3.43

Cloned(Commentary)

Cloned (Comment) Organism
variants of BphI expressed in recombinant Escherichia coli BL21, lambdaDE3 cells Paraburkholderia xenovorans

Protein Variants

Protein Variants Comment Organism
L87N mutant created by site-specific mutagenesis Paraburkholderia xenovorans
L87N/Y290F exhibits stereoselectivity opposite to that of the wild type and alkyl chain length of the aldehyde does not affect stereochemical control Paraburkholderia xenovorans
L87W mutant created by site-specific mutagenesis Paraburkholderia xenovorans
L87W/Y290F exhibits stereoselectivity opposite to that of the wild type and alkyl chain length of the aldehyde does not affect stereochemical control Paraburkholderia xenovorans
additional information mutants created by site-specific mutagenesis Paraburkholderia xenovorans
Y290F mutant is able to completely degrade racemic mixtures of 4-hydroxy-2-oxoacids i.e. is not stereospecific Paraburkholderia xenovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
 (4S)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.013
-
 (4R)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.089
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.21
-
 oxaloacetate L87N variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.435
-
 (4R)-4-hydroxy-2-oxopentanoate L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.757
-
 (4R)-4-hydroxy-2-oxopentanoate L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.84
-
 oxaloacetate L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.92
-
 oxaloacetate Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
6.2
-
 oxaloacetate L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
7.4
-
 oxaloacetate wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
13
-
 pyruvate wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
20
-
 pyruvate Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
24.4
-
 propionaldehyde L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
30.2
-
 pyruvate L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
38.2
-
 pyruvate L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
54.9
-
 propionaldehyde L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
64.2
-
 acetaldehyde wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
70.7
-
 acetaldehyde L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
80.5
-
 pentaldehyde Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
81
-
 acetaldehyde Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
81.2
-
 propionaldehyde Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
86.3
-
 acetaldehyde L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
104
-
 Butyraldehyde Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
136
-
 propionaldehyde wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans

Organism

Organism UniProt Comment Textmining
Paraburkholderia xenovorans
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(4R)-4-hydroxy-2-oxohexanoate reaction catalyzed by BphI variants Y290F, L87N/Y290F and L87W/Y290F Paraburkholderia xenovorans pyruvate + propionaldehyde
-
 ?
(4R)-4-hydroxy-2-oxopentanoate
-
 Paraburkholderia xenovorans pyruvate + acetaldehyde
-
 ?
(4S)-4-hydroxy-2-oxohexanoate double variants L87N/Y290F and L87W/Y290F inactive toward (4S)-4-hydroxy-2-oxohexanoate Paraburkholderia xenovorans pyruvate + propionaldehyde
-
 ?
(4S)-4-hydroxy-2-oxopentanoate
-
 Paraburkholderia xenovorans pyruvate + acetaldehyde
-
 ?
Oxaloacetate oxaloacetate decarboxylation is a secondary reaction of pyruvate aldolases Paraburkholderia xenovorans CO2 + pyruvate
-
 ?
pyruvate + acetaldehyde
-
 Paraburkholderia xenovorans (4R)-4-hydroxy-2-oxopentanoate
-
 ?
pyruvate + acetaldehyde
-
 Paraburkholderia xenovorans (4S)-4-hydroxy-2-oxopentanoate
-
 ?
pyruvate + butyraldehyde
-
 Paraburkholderia xenovorans ?
-
 ?
pyruvate + pentaldehyde
-
 Paraburkholderia xenovorans ?
-
 ?
pyruvate + propionaldehyde
-
 Paraburkholderia xenovorans 4-hydroxy-2-oxohexanoate
-
 ?

Synonyms

Synonyms Comment Organism
BphI
-
 Paraburkholderia xenovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.043
-
 (4R)-4-hydroxy-2-oxopentanoate L87W/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.05
-
 oxaloacetate L87N variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.08
-
 oxaloacetate L87N/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.098
-
 (4R)-4-hydroxy-2-oxopentanoate L87N/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.1
-
 propionaldehyde L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.11
-
 pyruvate L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.131
-
 (4R)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.132
-
 (4S)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.14
-
 propionaldehyde L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.14
-
 pentaldehyde Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.16
-
 pyruvate L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.16
-
 propionaldehyde Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.16
-
 Butyraldehyde Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.19
-
 pyruvate Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.2
-
 acetaldehyde Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.27
-
 oxaloacetate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.3
-
 acetaldehyde L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.4
-
 oxaloacetate L87W/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.59
-
 acetaldehyde L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.86
-
 acetaldehyde wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
1.2
-
 pyruvate wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
1.79
-
 propionaldehyde wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
4
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
16.3
-
 oxaloacetate wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00004
-
 pentaldehyde L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.000051
-
 pentaldehyde L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.00016
-
 Butyraldehyde L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.00031
-
 Butyraldehyde L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.00047
-
 pentaldehyde wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0015
-
 Butyraldehyde Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0017
-
 pentaldehyde Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0018
-
 propionaldehyde L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.002
-
 propionaldehyde Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.00219
-
 Butyraldehyde wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0025
-
 acetaldehyde Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0036
-
 pyruvate L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0043
-
 acetaldehyde L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0043
-
 pyruvate L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0057
-
 propionaldehyde L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0058
-
 oxaloacetate L87W variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0069
-
 acetaldehyde L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0084
-
 pyruvate Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0132
-
 propionaldehyde wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0134
-
 acetaldehyde wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.064
-
 oxaloacetate L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0915
-
 pyruvate wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.098
-
 oxaloacetate L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.1
-
 (4R)-4-hydroxy-2-oxopentanoate L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.13
-
 (4R)-4-hydroxy-2-oxopentanoate L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.254
-
 oxaloacetate L87N variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.53
-
 oxaloacetate Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
2.2
-
 oxaloacetate wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
10.4
-
 (4R)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
12.6
-
 (4S)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
44.7
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans