BRENDA - Enzyme Database
show all sequences of 4.1.3.43

Investigating the molecular determinants for substrate channeling in BphI-BphJ, an aldolase-dehydrogenase complex from the polychlorinated biphenyls degradation pathway

Carere, J.; Baker, P.; Seah, S.Y.K.; Biochemistry 50, 8407-8416 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
bphI and bphJ cloned into the plasmids pBTL4-T7 and pET28a
Paraburkholderia xenovorans
Engineering
Amino acid exchange
Commentary
Organism
G322A
channels acetaldehyde with similar efficiency to wild-type, 30% lowered efficiency of isobutyraldehyde channeling
Paraburkholderia xenovorans
G322F
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G322L
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G323A
channels acetaldehyde with similar efficiency to wild-type, unable to channel isobutyraldehyde
Paraburkholderia xenovorans
G323F
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G323L
63% channeling efficiency for acetaldehyde, unable to channel propionaldehyde
Paraburkholderia xenovorans
H20A
reduction of acetaldehyde and propionaldehyde channeling by more than 70%
Paraburkholderia xenovorans
L89A
reduction in channeling efficiency of 30% for all aldehydes tested
Paraburkholderia xenovorans
additional information
all mutations created by site specific mutagenesis
Paraburkholderia xenovorans
Y290F
reduction in channeling efficiency of 30% for all aldehydes tested
Paraburkholderia xenovorans
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.053
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.06
-
4-hydroxy-2-oxopentanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.081
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.089
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.09
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.117
-
4-hydroxy-2-oxohexanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.13
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.14
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.158
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.16
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.18
-
4-hydroxy-2-oxopentanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.216
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.3
-
4-hydroxy-2-oxohexanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.35
-
4-hydroxy-2-oxohexanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.36
-
4-hydroxy-2-oxohexanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.37
-
4-hydroxy-2-oxohexanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.52
-
4-hydroxy-2-oxohexanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.94
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.1
-
4-hydroxy-2-oxohexanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Paraburkholderia xenovorans
-
-
-
Purification (Commentary)
Commentary
Organism
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
Paraburkholderia xenovorans
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-hydroxy-2-oxohexanoate
4-hydroxy-2-oxohexanoate i.e. HOHA
721651
Paraburkholderia xenovorans
propionaldehyde + pyruvate
-
-
-
?
4-hydroxy-2-oxopentanoate
4-hydroxy-2-oxopentanoate i.e. HOPA
721651
Paraburkholderia xenovorans
acetaldehyde + pyruvate
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.1
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.27
-
4-hydroxy-2-oxohexanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.286
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.34
-
4-hydroxy-2-oxopentanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.45
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.46
-
4-hydroxy-2-oxohexanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.64
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.72
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.79
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.94
-
4-hydroxy-2-oxohexanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.05
-
4-hydroxy-2-oxohexanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.06
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.09
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.3
-
4-hydroxy-2-oxohexanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.9
-
4-hydroxy-2-oxohexanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
2.4
-
4-hydroxy-2-oxopentanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
3.9
-
4-hydroxy-2-oxohexanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
4.07
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
4.3
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
Cloned(Commentary) (protein specific)
Commentary
Organism
bphI and bphJ cloned into the plasmids pBTL4-T7 and pET28a
Paraburkholderia xenovorans
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
G322A
channels acetaldehyde with similar efficiency to wild-type, 30% lowered efficiency of isobutyraldehyde channeling
Paraburkholderia xenovorans
G322F
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G322L
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G323A
channels acetaldehyde with similar efficiency to wild-type, unable to channel isobutyraldehyde
Paraburkholderia xenovorans
G323F
unable to channel either acetaldehyde or propionaldehyde
Paraburkholderia xenovorans
G323L
63% channeling efficiency for acetaldehyde, unable to channel propionaldehyde
Paraburkholderia xenovorans
H20A
reduction of acetaldehyde and propionaldehyde channeling by more than 70%
Paraburkholderia xenovorans
L89A
reduction in channeling efficiency of 30% for all aldehydes tested
Paraburkholderia xenovorans
additional information
all mutations created by site specific mutagenesis
Paraburkholderia xenovorans
Y290F
reduction in channeling efficiency of 30% for all aldehydes tested
Paraburkholderia xenovorans
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.053
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.06
-
4-hydroxy-2-oxopentanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.081
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.089
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.09
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.117
-
4-hydroxy-2-oxohexanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.13
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.14
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.158
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.16
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.18
-
4-hydroxy-2-oxopentanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.216
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.3
-
4-hydroxy-2-oxohexanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.35
-
4-hydroxy-2-oxohexanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.36
-
4-hydroxy-2-oxohexanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.37
-
4-hydroxy-2-oxohexanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.52
-
4-hydroxy-2-oxohexanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.94
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.1
-
4-hydroxy-2-oxohexanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
Purification (Commentary) (protein specific)
Commentary
Organism
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
Paraburkholderia xenovorans
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-hydroxy-2-oxohexanoate
4-hydroxy-2-oxohexanoate i.e. HOHA
721651
Paraburkholderia xenovorans
propionaldehyde + pyruvate
-
-
-
?
4-hydroxy-2-oxopentanoate
4-hydroxy-2-oxopentanoate i.e. HOPA
721651
Paraburkholderia xenovorans
acetaldehyde + pyruvate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.1
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.27
-
4-hydroxy-2-oxohexanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.286
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.34
-
4-hydroxy-2-oxopentanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.45
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.46
-
4-hydroxy-2-oxohexanoate
G323A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.64
-
4-hydroxy-2-oxopentanoate
G323L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
0.72
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.79
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C
Paraburkholderia xenovorans
0.94
-
4-hydroxy-2-oxohexanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.05
-
4-hydroxy-2-oxohexanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.06
-
4-hydroxy-2-oxopentanoate
G322F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.09
-
4-hydroxy-2-oxopentanoate
G322L mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.3
-
4-hydroxy-2-oxohexanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
1.9
-
4-hydroxy-2-oxohexanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
2.4
-
4-hydroxy-2-oxopentanoate
G322A mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
3.9
-
4-hydroxy-2-oxohexanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
4.07
-
4-hydroxy-2-oxopentanoate
wild-type protein, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
4.3
-
4-hydroxy-2-oxopentanoate
G323F mutant, steady state, pH 8.0, 25°C
Paraburkholderia xenovorans
General Information
General Information
Commentary
Organism
metabolism
enzyme is part of the BphI-BphJ-complex, an aldolase-dehydrogenase complex from the polychlorinated biphenyl degradation pathway
Paraburkholderia xenovorans
General Information (protein specific)
General Information
Commentary
Organism
metabolism
enzyme is part of the BphI-BphJ-complex, an aldolase-dehydrogenase complex from the polychlorinated biphenyl degradation pathway
Paraburkholderia xenovorans
Other publictions for EC 4.1.3.43
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
721659
Baker
Protein-protein interactions a ...
Thermus thermophilus
Biochemistry
51
1942-1952
2012
-
-
1
-
1
-
-
16
-
-
7
-
-
4
-
-
1
-
-
-
-
-
4
1
-
-
1
15
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
16
-
-
7
-
-
-
-
1
-
-
-
-
4
1
-
-
1
15
-
-
-
-
-
1
1
-
15
15
722458
Baker
Rational design of stereoselec ...
Paraburkholderia xenovorans
J. Am. Chem. Soc.
134
507-513
2012
-
-
1
-
6
-
-
24
-
-
-
-
-
1
-
-
-
-
-
-
-
-
10
-
-
-
-
24
-
-
-
-
-
-
-
-
-
1
-
-
6
-
-
-
-
24
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
-
24
-
-
-
-
-
-
-
-
31
31
714244
Baker
Probing the molecular basis of ...
Paraburkholderia xenovorans
Biochemistry
50
3559-3569
2011
-
-
1
-
7
-
-
20
-
-
-
-
-
1
-
-
-
-
-
-
-
-
9
-
-
-
-
21
-
-
-
-
-
-
-
-
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1
-
-
7
-
-
-
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20
-
-
-
-
-
-
-
-
-
-
-
-
9
-
-
-
-
21
-
-
-
-
-
1
1
-
31
31
721651
Carere
Investigating the molecular de ...
Paraburkholderia xenovorans
Biochemistry
50
8407-8416
2011
-
-
1
-
10
-
-
19
-
-
-
-
-
1
-
-
1
-
-
-
-
-
2
-
-
-
-
19
-
-
-
-
-
-
-
-
-
1
-
-
10
-
-
-
-
19
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
19
-
-
-
-
-
1
1
-
-
-
702377
Wang
Comparison of two metal-depend ...
Paraburkholderia xenovorans
Biochemistry
49
3774-3782
2010
-
-
-
-
-
-
5
5
-
1
-
-
-
1
-
-
-
-
-
-
-
-
6
-
-
-
-
4
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
5
6
5
-
1
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
4
-
-
-
-
-
-
-
-
6
6
721615
Baker
Characterization of an aldolas ...
Paraburkholderia xenovorans
Biochemistry
48
6551-6558
2009
1
-
1
-
-
-
6
5
-
-
3
1
-
1
-
-
1
-
-
-
-
1
5
1
-
-
-
5
-
-
-
-
1
-
-
1
-
1
-
-
-
-
-
6
1
5
-
-
3
1
-
-
-
1
-
-
-
1
5
1
-
-
-
5
-
-
-
-
-
1
1
-
5
5