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Literature summary for 4.1.3.43 extracted from

  • Baker, P.; Carere, J.; Seah, S.Y.
    Probing the molecular basis of substrate specificity, stereospecificity, and catalysis in the class II pyruvate aldolase, BphI (2011), Biochemistry, 50, 3559-3569.
    View publication on PubMed
Search for more literature for 4.1.3.43

Cloned(Commentary)

Cloned (Comment) Organism
bphI previous to mutagenesis cloned into plasmid pBTL4-T7, Escherichia coli Paraburkholderia xenovorans

Protein Variants

Protein Variants Comment Organism
H20A mutant generated by site-directed mutagenesis, pH profiles show the dependence of enzyme activity on hydroxide concentration Paraburkholderia xenovorans
H20S mutant generated by site-directed mutagenesis, pH profiles show the dependence of enzyme activity on hydroxide concentration Paraburkholderia xenovorans
L87A mutant generated by site-directed mutagenesis, variant exhibits a 40fold preference for propionaldehyde over acetaldehyde in contrast to wild type with similar specificities for acetaldehyde and propionaldehyde Paraburkholderia xenovorans
L89A mutant generated by site-directed mutagenesis, specificity constant of this variant in the aldol addition reaction using pentaldehyde is increased approx. 50fold, making it more catalytically efficient for pentaldehyde utilization compared to the wild-type utilization of the natural substrate, acetaldehyde Paraburkholderia xenovorans
R16A mutant generated by site-directed mutagenesis, no detectable aldol cleavage and pyruvate R-proton exchange, supporting the role of Arg-16 in stabilizing a pyruvate enolate intermediate Paraburkholderia xenovorans
Y290F mutant generated by site-directed mutagenesis, mutation results in a loss of stereochemical control as the variant is able to utilize substrates with R and S configurations at C4 with similar kinetic parameters Paraburkholderia xenovorans
Y290S mutant generated by site-directed mutagenesis, mutation results in a loss of stereochemical control as the variant is able to utilize substrates with R and S configurations at C4 with similar kinetic parameters Paraburkholderia xenovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information value not detectable for R16A variant of BphI Paraburkholderia xenovorans
0.012
-
 (4R)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.013
-
 (4S)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.013
-
 (4R)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.013
-
 (4S)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.089
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.222
-
 (4S)-4-hydroxy-2-oxopentanoate H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.228
-
 (4S)-4-hydroxy-2-oxopentanoate H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
11
-
 pyruvate L89A variant of 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
16.8
-
 propionaldehyde L87A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
20.2
-
 pyruvate L87A variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
24.8
-
 pentaldehyde L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
33.3
-
 pentaldehyde L87A/L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
40.2
-
 propionaldehyde L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
49.2
-
 Butyraldehyde L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
64.2
-
 acetaldehyde wild-type BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
80.5
-
 acetaldehyde L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
136
-
 propionaldehyde wild-type BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
289
-
 Butyraldehyde L87A/L89A variant of BphI, app. Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
323
-
 Butyraldehyde L87A variant of BphI, app. Km-value, 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-oxopentanoate
-
 Paraburkholderia xenovorans pyruvate + acetaldehyde
-
 ?
(4S)-4-hydroxy-2-oxopentanoate
-
 Paraburkholderia xenovorans pyruvate + acetaldehyde
-
 ?
pyruvate + acetaldehyde
-
 Paraburkholderia xenovorans (4R)-4-hydroxy-2-oxopentanoate
-
 ?
pyruvate + acetaldehyde
-
 Paraburkholderia xenovorans (4S)-4-hydroxy-2-oxopentanoate
-
 ?
pyruvate + butyraldehyde
-
 Paraburkholderia xenovorans ?
-
 ?
pyruvate + hexaldehyde
-
 Paraburkholderia xenovorans ?
-
 ?
pyruvate + pentaldehyde
-
 Paraburkholderia xenovorans ?
-
 ?
pyruvate + propionaldehyde reaction catalyzed by BphI variants Y290F and Y290S Paraburkholderia xenovorans (4R)-4-hydroxy-2-oxohexanoate
-
 ?
pyruvate + propionaldehyde
-
 Paraburkholderia xenovorans (4S)-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
additional information
-
 additional information value not detectable for R16A variant of BphI Paraburkholderia xenovorans
0.089
-
 (4S)-4-hydroxy-2-oxopentanoate H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.099
-
 (4S)-4-hydroxy-2-oxopentanoate H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.13
-
 pentaldehyde L87A/L89A variant of BphI, app. kcat-value, 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
0.17
-
 (4R)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.171
-
 (4S)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.26
-
 Butyraldehyde L87A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.29
-
 propionaldehyde L87A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.3
-
 Butyraldehyde L87A/L89A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.32
-
 pyruvate L87A variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.58
-
 pentaldehyde L89A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.63
-
 propionaldehyde L89A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.64
-
 Butyraldehyde L89A variant of BphI, app. kcat-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.64
-
 pyruvate L89A 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, pH 8.0, 25°C Paraburkholderia xenovorans
0.94
-
 acetaldehyde L89A variant of BphI, app. kcat-value, 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, pH 8.0, 25°C Paraburkholderia xenovorans
4.07
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans

General Information

General Information Comment Organism
metabolism enzyme found in the polychlorinated biphenyls (PCBs) degradation pathway Paraburkholderia xenovorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
 additional information value not detectable for R16A variant of BphI Paraburkholderia xenovorans
0.00001
-
 hexaldehyde L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00001
-
 hexaldehyde wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0001
-
 hexaldehyde L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00013
-
 hexaldehyde L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00033
-
 pentaldehyde L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00042
-
 acetaldehyde L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00047
-
 pentaldehyde wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00049
-
 acetaldehyde L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00079
-
 Butyraldehyde L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00104
-
 Butyraldehyde L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0022
-
 Butyraldehyde wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0038
-
 pentaldehyde L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.00673
-
 propionaldehyde L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0117
-
 acetaldehyde L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.012
-
 (4S)-4-hydroxy-2-oxopentanoate R16K variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0129
-
 Butyraldehyde L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0132
-
 propionaldehyde wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0134
-
 acetaldehyde wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0156
-
 propionaldehyde L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0159
-
 pyruvate L87A variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.0175
-
 propionaldehyde L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0233
-
 pentaldehyde L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C Paraburkholderia xenovorans
0.0587
-
 pyruvate L89A 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
0.34
-
 (4S)-4-hydroxy-2-oxopentanoate H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
0.45
-
 (4S)-4-hydroxy-2-oxopentanoate H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
10.4
-
 (4R)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
12.6
-
 (4S)-4-hydroxy-2-oxopentanoate Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
13.4
-
 (4S)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
14.1
-
 (4R)-4-hydroxy-2-oxopentanoate Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans
44.7
-
 (4S)-4-hydroxy-2-oxopentanoate wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C Paraburkholderia xenovorans