Any feedback?
Information on EC 4.1.3.43 - 4-hydroxy-2-oxohexanoate aldolase and Organism(s) Paraburkholderia xenovorans and UniProt Accession P51015
for references in articles please use BRENDA:EC4.1.3.43Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
4.1.3.43 4-hydroxy-2-oxohexanoate aldolaseIUBMB Comments
Requires Mn2+ for maximal activity [1,2]. The enzymes from the bacteria Burkholderia xenovorans and Thermus thermophilus also perform the reaction of EC 4.1.3.39, 4-hydroxy-2-oxovalerate aldolase [1,2,6]. The enzyme forms a bifunctional complex with EC 1.2.1.87, propanal dehydrogenase (CoA-propanoylating), with a tight channel connecting the two subunits [3,4,6].
Specify your search results
The taxonomic range for the selected organisms is: Paraburkholderia xenovorans
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
tthb246, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
BphI
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
BphI exhibits a compulsory order mechanism, with pyruvate binding first
PATHWAY SOURCE
PATHWAYS
-
-, -, -
SYSTEMATIC NAME
IUBMB Comments
(S)-4-hydroxy-2-oxohexanoate pyruvate-lyase (propanal-forming)
Requires Mn2+ for maximal activity [1,2]. The enzymes from the bacteria Burkholderia xenovorans and Thermus thermophilus also perform the reaction of EC 4.1.3.39, 4-hydroxy-2-oxovalerate aldolase [1,2,6]. The enzyme forms a bifunctional complex with EC 1.2.1.87, propanal dehydrogenase (CoA-propanoylating), with a tight channel connecting the two subunits [3,4,6].
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(4R)-4-hydroxy-2-oxohexanoate
pyruvate + propionaldehyde
-
reaction catalyzed by BphI variants Y290F, L87N/Y290F and L87W/Y290F
-
-
?
(4R)-4-hydroxy-2-oxopentanoate
pyruvate + acetaldehyde
-
-
-
-
?
(4S)-4-hydroxy-2-oxohexanoate
pyruvate + propionaldehyde
-
double variants L87N/Y290F and L87W/Y290F inactive toward (4S)-4-hydroxy-2-oxohexanoate
-
-
?
(4S)-4-hydroxy-2-oxopentanoate
pyruvate + acetaldehyde
-
-
-
-
?
4-hydroxy-2-oxohexanoate
propionaldehyde + pyruvate
-
4-hydroxy-2-oxohexanoate i.e. HOHA
-
-
?
4-hydroxy-2-oxohexanoate
pyruvate + propionaldehyde
-
-
-
?
4-hydroxy-2-oxopentanoate
acetaldehyde + pyruvate
-
4-hydroxy-2-oxopentanoate i.e. HOPA
-
-
?
Oxaloacetate
CO2 + pyruvate
-
oxaloacetate decarboxylation is a secondary reaction of pyruvate aldolases
-
-
?
pyruvate + acetaldehyde
(4R)-4-hydroxy-2-oxopentanoate
-
-
-
-
?
pyruvate + acetaldehyde
(4S)-4-hydroxy-2-oxopentanoate
-
-
-
-
?
pyruvate + propionaldehyde
(4R)-4-hydroxy-2-oxohexanoate
-
reaction catalyzed by BphI variants Y290F and Y290S
-
-
?
pyruvate + propionaldehyde
(4S)-4-hydroxy-2-oxohexanoate
-
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Cd2+
56.0% activity at 0.1 mM and 5.4% activity at 1 mM chloride salt
Co2+
19.3% activity at 0.1 mM and 18.3% activity at 1 mM chloride salt
Mg2+
3.4% activity at 0.1 mM and 14.8% activity at 1 mM chloride salt
Ni2+
2.6% activity at 0.1 mM and 17.5% activity at 1 mM chloride salt
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
124.6
glycolaldehyde
-
app. Km-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.012
(4R)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.013
(4R)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.013
(4R)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.435
(4R)-4-hydroxy-2-oxopentanoate
-
L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.757
(4R)-4-hydroxy-2-oxopentanoate
-
L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.013
(4S)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.013
(4S)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.013
(4S)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.089
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.089
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.222
(4S)-4-hydroxy-2-oxopentanoate
-
H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.228
(4S)-4-hydroxy-2-oxopentanoate
-
H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.117
4-hydroxy-2-oxohexanoate
-
wild-type protein, steady state, pH 8.0, 25°C
0.3
4-hydroxy-2-oxohexanoate
-
G322F mutant, steady state, pH 8.0, 25°C
0.35
4-hydroxy-2-oxohexanoate
-
G322L mutant, steady state, pH 8.0, 25°C
0.36
4-hydroxy-2-oxohexanoate
-
G323A mutant, steady state, pH 8.0, 25°C
0.37
4-hydroxy-2-oxohexanoate
-
G323L mutant, steady state, pH 8.0, 25°C
0.52
4-hydroxy-2-oxohexanoate
-
G322A mutant, steady state, pH 8.0, 25°C
1.1
4-hydroxy-2-oxohexanoate
-
G323F mutant, steady state, pH 8.0, 25°C
0.053
4-hydroxy-2-oxopentanoate
-
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.06
4-hydroxy-2-oxopentanoate
-
G323A mutant, steady state, pH 8.0, 25°C
0.081
4-hydroxy-2-oxopentanoate
-
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.089
4-hydroxy-2-oxopentanoate
-
wild-type protein, steady state, pH 8.0, 25°C
0.09
4-hydroxy-2-oxopentanoate
-
G322F mutant, steady state, pH 8.0, 25°C
0.13
4-hydroxy-2-oxopentanoate
-
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.14
4-hydroxy-2-oxopentanoate
-
G322L mutant, steady state, pH 8.0, 25°C
0.158
4-hydroxy-2-oxopentanoate
-
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.16
4-hydroxy-2-oxopentanoate
-
G323L mutant, steady state, pH 8.0, 25°C
0.18
4-hydroxy-2-oxopentanoate
-
G322A mutant, steady state, pH 8.0, 25°C
0.216
4-hydroxy-2-oxopentanoate
-
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.94
4-hydroxy-2-oxopentanoate
-
G323F mutant, steady state, pH 8.0, 25°C
64.2
acetaldehyde
-
wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
64.3
acetaldehyde
-
app. Km-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
67
acetaldehyde
-
steady-state kinetic parameter, pH 8.0 and 25°C
70.7
acetaldehyde
-
L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
80.5
acetaldehyde
-
L89A variant of BphI, app. Km-value, pH 8.0, 25°C
81
acetaldehyde
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
86.3
acetaldehyde
-
L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
49.2
Butyraldehyde
-
L89A variant of BphI, app. Km-value, pH 8.0, 25°C
104
Butyraldehyde
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
289
Butyraldehyde
-
L87A/L89A variant of BphI, app. Km-value, pH 8.0, 25°C
323
Butyraldehyde
-
L87A variant of BphI, app. Km-value, pH 8.0, 25°C
0.21
oxaloacetate
-
L87N variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.84
oxaloacetate
-
L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.92
oxaloacetate
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
6.2
oxaloacetate
-
L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
7.4
oxaloacetate
-
wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
24.8
pentaldehyde
-
L89A variant of BphI, app. Km-value, pH 8.0, 25°C
33.3
pentaldehyde
-
L87A/L89A variant of BphI, app. Km-value, pH 8.0, 25°C
80.5
pentaldehyde
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
16.8
propionaldehyde
-
L87A variant of BphI, app. Km-value, pH 8.0, 25°C
24.4
propionaldehyde
-
L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
40.2
propionaldehyde
-
L89A variant of BphI, app. Km-value, pH 8.0, 25°C
54.9
propionaldehyde
-
L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
81.2
propionaldehyde
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
135.9
propionaldehyde
-
app. Km-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
136
propionaldehyde
-
wild-type BphI, app. Km-value, pH 8.0, 25°C
136
propionaldehyde
-
wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
11
pyruvate
-
L89A variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
13
pyruvate
-
wild-type BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
20
pyruvate
-
Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
20.2
pyruvate
-
L87A variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
30.2
pyruvate
-
L87N/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
38.2
pyruvate
-
L87W/Y290F variant of BphI, app. Km-value, steady-state kinetic parameter, pH 8.0, 25°C
additional information
additional information
presence of NAD+, NADP+, NADH or CoA leads to an at least 5fold reduction in the Km for 4-hydroxy-2-oxopentanoate
-
additional information
additional information
-
value not detectable for R16A variant of BphI
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.4
glycolaldehyde
-
app. kcat-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.043
(4R)-4-hydroxy-2-oxopentanoate
-
L87W/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.098
(4R)-4-hydroxy-2-oxopentanoate
-
L87N/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.131
(4R)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.17
(4R)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.089
(4S)-4-hydroxy-2-oxopentanoate
-
H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.099
(4S)-4-hydroxy-2-oxopentanoate
-
H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.132
(4S)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.171
(4S)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
4
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C
4.07
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.27
4-hydroxy-2-oxohexanoate
-
G323L mutant, steady state, pH 8.0, 25°C
0.46
4-hydroxy-2-oxohexanoate
-
G323A mutant, steady state, pH 8.0, 25°C
0.94
4-hydroxy-2-oxohexanoate
-
G322L mutant, steady state, pH 8.0, 25°C
1.05
4-hydroxy-2-oxohexanoate
-
G322F mutant, steady state, pH 8.0, 25°C
1.3
4-hydroxy-2-oxohexanoate
-
G323F mutant, steady state, pH 8.0, 25°C
1.9
4-hydroxy-2-oxohexanoate
-
G322A mutant, steady state, pH 8.0, 25°C
3.9
4-hydroxy-2-oxohexanoate
-
wild-type protein, steady state, pH 8.0, 25°C
0.1
4-hydroxy-2-oxopentanoate
-
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.286
4-hydroxy-2-oxopentanoate
-
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.34
4-hydroxy-2-oxopentanoate
-
G323A mutant, steady state, pH 8.0, 25°C
0.45
4-hydroxy-2-oxopentanoate
-
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.64
4-hydroxy-2-oxopentanoate
-
G323L mutant, steady state, pH 8.0, 25°C
0.72
4-hydroxy-2-oxopentanoate
-
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
0.79
4-hydroxy-2-oxopentanoate
-
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
1.06
4-hydroxy-2-oxopentanoate
-
G322F mutant, steady state, pH 8.0, 25°C
1.09
4-hydroxy-2-oxopentanoate
-
G322L mutant, steady state, pH 8.0, 25°C
2.4
4-hydroxy-2-oxopentanoate
-
G322A mutant, steady state, pH 8.0, 25°C
4.07
4-hydroxy-2-oxopentanoate
-
wild-type protein, steady state, pH 8.0, 25°C
4.3
4-hydroxy-2-oxopentanoate
-
G323F mutant, steady state, pH 8.0, 25°C
0.2
acetaldehyde
-
Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.3
acetaldehyde
-
L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.59
acetaldehyde
-
L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.86
acetaldehyde
-
app. kcat-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.86
acetaldehyde
-
wild-type BphI, app. kcat-value, pH 8.0, 25°C
0.86
acetaldehyde
-
wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.94
acetaldehyde
-
L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.16
Butyraldehyde
-
Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.26
Butyraldehyde
-
L87A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.3
Butyraldehyde
-
L87A/L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.64
Butyraldehyde
-
L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.05
oxaloacetate
-
L87N variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.08
oxaloacetate
-
L87N/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.27
oxaloacetate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.4
oxaloacetate
-
L87W/Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
16.3
oxaloacetate
-
wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.13
pentaldehyde
-
L87A/L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.14
pentaldehyde
-
Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.58
pentaldehyde
-
L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.1
propionaldehyde
-
L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.14
propionaldehyde
-
L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.16
propionaldehyde
-
Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.29
propionaldehyde
-
L87A variant of BphI, app. kcat-value, pH 8.0, 25°C
0.63
propionaldehyde
-
L89A variant of BphI, app. kcat-value, pH 8.0, 25°C
1.79
propionaldehyde
-
app. kcat-value with pyruvate as carbonyl donor, pH 8.0 and 25°C
1.79
propionaldehyde
-
wild-type BphI, app. kcat-value, pH 8.0, 25°C
1.79
propionaldehyde
-
wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.11
pyruvate
-
L87N/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.16
pyruvate
-
L87W/Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.19
pyruvate
-
Y290F variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.32
pyruvate
-
L87A variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
0.64
pyruvate
-
L89A variant of BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
1.04
pyruvate
-
steady-state kinetic parameter, with acetaldehyde as aldehyde donor, pH 8.0 and 25°C
1.2
pyruvate
-
wild-type BphI, app. kcat-value, steady-state kinetic parameter, pH 8.0, 25°C
additional information
additional information
2 and 5fold increases in kcat observed when BphI reaction is conducted in the presence of NAD+ and NADH
-
additional information
additional information
-
value not detectable for R16A variant of BphI
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0037
glycolaldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.0005
Isobutyraldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
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
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
10.4
(4R)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
10.4
(4R)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
14.1
(4R)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.012
(4S)-4-hydroxy-2-oxopentanoate
-
R16K variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.34
(4S)-4-hydroxy-2-oxopentanoate
-
H20S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.45
(4S)-4-hydroxy-2-oxopentanoate
-
H20A variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
12.6
(4S)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
12.6
(4S)-4-hydroxy-2-oxopentanoate
-
Y290F variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
13.4
(4S)-4-hydroxy-2-oxopentanoate
-
Y290S variant of BphI, steady-state kinetic parameter, pH 8.0, 25°C
44.7
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
44.7
(4S)-4-hydroxy-2-oxopentanoate
-
wild-type BphI, steady-state kinetic parameter, pH 8.0, 25°C
0.00042
acetaldehyde
-
L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00049
acetaldehyde
-
L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0025
acetaldehyde
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0043
acetaldehyde
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0069
acetaldehyde
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0117
acetaldehyde
-
L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0134
acetaldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.0134
acetaldehyde
-
wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0134
acetaldehyde
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00016
Butyraldehyde
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00031
Butyraldehyde
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00079
Butyraldehyde
-
L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00104
Butyraldehyde
-
L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0015
Butyraldehyde
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00219
Butyraldehyde
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0022
Butyraldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.0022
Butyraldehyde
-
wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0129
Butyraldehyde
-
L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00001
hexaldehyde
-
L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00001
hexaldehyde
-
wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0001
hexaldehyde
-
L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00013
hexaldehyde
-
L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0058
oxaloacetate
-
L87W variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.064
oxaloacetate
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.098
oxaloacetate
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.254
oxaloacetate
-
L87N variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.53
oxaloacetate
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
2.2
oxaloacetate
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00004
pentaldehyde
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.000051
pentaldehyde
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00033
pentaldehyde
-
L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00047
pentaldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.00047
pentaldehyde
-
wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C
0.00047
pentaldehyde
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0017
pentaldehyde
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0038
pentaldehyde
-
L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0233
pentaldehyde
-
L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0018
propionaldehyde
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.002
propionaldehyde
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0057
propionaldehyde
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.00673
propionaldehyde
-
L87A/L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0132
propionaldehyde
-
kcat(app)/Km(app) with pyruvate as carbonyl donor, pH 8.0 and 25°C
0.0132
propionaldehyde
-
wild-type BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0132
propionaldehyde
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0156
propionaldehyde
-
L89A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0175
propionaldehyde
-
L87A variant of BphI, app. kcat/Km-value, pH 8.0, 25°C
0.0036
pyruvate
-
L87N/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0043
pyruvate
-
L87W/Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0084
pyruvate
-
Y290F variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0159
pyruvate
-
L87A variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0587
pyruvate
-
L89A variant of BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
0.0915
pyruvate
-
wild-type BphI, app. kcat/Km-value, steady-state kinetic parameter, pH 8.0, 25°C
additional information
additional information
highest catalytic efficiency of BphI occurs when NADH is present, which is 25fold higher than the value obtained without nucleotides
-
additional information
additional information
-
value not detectable for R16A variant of BphI
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
metabolism
enzyme forms a bifunctional complex with EC 1.2.1.87 (BphJ), propanal dehydrogenase (CoA-propanoylating) and is part of the polychlorinated biphenyl degradation pathway
metabolism
-
enzyme found in the polychlorinated biphenyls (PCBs) degradation pathway
metabolism
-
enzyme is part of the BphI-BphJ-complex, an aldolase-dehydrogenase complex from the polychlorinated biphenyl degradation pathway
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
140000
native molecular mass of the purified BphI-BphJ-complex, estimated by gel filtration
32000
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI
37000
SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
heterotetramer
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
G322A
-
channels acetaldehyde with similar efficiency to wild-type, 30% lowered efficiency of isobutyraldehyde channeling
G323A
-
channels acetaldehyde with similar efficiency to wild-type, unable to channel isobutyraldehyde
G323L
-
63% channeling efficiency for acetaldehyde, unable to channel propionaldehyde
H20A
H20S
-
mutant generated by site-directed mutagenesis, pH profiles show the dependence of enzyme activity on hydroxide concentration
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
L87N/Y290F
-
exhibits stereoselectivity opposite to that of the wild type and alkyl chain length of the aldehyde does not affect stereochemical control
L87W/Y290F
-
exhibits stereoselectivity opposite to that of the wild type and alkyl chain length of the aldehyde does not affect stereochemical control
L89A
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
Y290F
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
additional information
H20A
-
mutant generated by site-directed mutagenesis, pH profiles show the dependence of enzyme activity on hydroxide concentration
H20A
-
reduction of acetaldehyde and propionaldehyde channeling by more than 70%
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
L89A
-
reduction in channeling efficiency of 30% for all aldehydes tested
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
Y290F
-
mutant is able to completely degrade racemic mixtures of 4-hydroxy-2-oxoacids i.e. is not stereospecific
Y290F
-
reduction in channeling efficiency of 30% for all aldehydes tested
additional information
-
all mutations created by site specific mutagenesis
additional information
-
mutants created by site-specific mutagenesis
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
purified enzyme can be stored at -80°C, without loss of activity for at least 12 months
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
BphI and BphJ form a stable complex as they bind and coelute from Ni2+-NTA column, after purification N-terminal histidine tag of BphJ is proteolytically cleaved by thrombin digestion
-
BphI and BphJ form a stable complex as they bind and coelute from Ni2+-NTA column, although only BphJ has the histidine tag. After purification, the N-terminal histidine tag of BphJ is proteolytically cleaved by thrombin digestion
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
bphI previous to mutagenesis cloned into plasmid pBTL4-T7, Escherichia coli
-
coexpression of bphI and bphJ in Escherichia coli using two compatible plasmids (pBTL4 and pET28a) yielded soluble proteins
variants of BphI expressed in recombinant Escherichia coli BL21, lambdaDE3 cells
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wang, W.; Baker, P.; Seah, S.Y.
Comparison of two metal-dependent pyruvate aldolases related by convergent evolution: substrate specificity, kinetic mechanism, and substrate channeling
Biochemistry
49
3774-3782
2010
Paraburkholderia xenovorans
Baker, P.; Carere, J.; Seah, S.Y.
Probing the molecular basis of substrate specificity, stereospecificity, and catalysis in the class II pyruvate aldolase, BphI
Biochemistry
50
3559-3569
2011
Paraburkholderia xenovorans
Baker, P.; Pan, D.; Carere, J.; Rossi, A.; Wang, W.; Seah, S.Y.K.
Characterization of an aldolase-dehydrogenase complex that exhibits substrate channeling in the polychlorinated biphenyls degradation pathway
Biochemistry
48
6551-6558
2009
Paraburkholderia xenovorans (P51015)
Carere, J.; Baker, P.; Seah, S.Y.K.
Investigating the molecular determinants for substrate channeling in BphI-BphJ, an aldolase-dehydrogenase complex from the polychlorinated biphenyls degradation pathway
Biochemistry
50
8407-8416
2011
Paraburkholderia xenovorans
EXTERNAL LINKS (specific for EC-Number 4.1.3.43)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
