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show all sequences of 2.8.3.18

Crystal structures of Acetobacter aceti succinyl-coenzyme A (CoA):acetate CoA-transferase reveal specificity determinants and illustrate the mechanism used by class I CoA-transferases

Mullins, E.A.; Kappock, T.J.; Biochemistry 51, 8422-8434 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli; gene aarC, sequence comparisons and phylogenetic analysis, expression of C-terminally His6-tagged wild-type and mutant enzymes
Acetobacter aceti
Crystallization (Commentary)
Crystallization
Organism
crystal structures of a C-terminally His6-tagged form of several wild-type and mutant complexes, including freeze-trapped acetylglutamyl anhydride and glutamyl-CoA thioester adducts. The latter shows the acetate product bound to an auxiliary site that is required for efficient carboxylate substrate recognition. Mutant E294A crystallizes in a closed complex containing dethiaacetyl-CoA, which adopts an unusual curled conformation. A model of the acetyl-CoA Michaelis complex reveals that the nucleophilic glutamate is held at a near-ideal angle for attack as the thioester oxygen is forced into an oxyanion hole composed of Gly388 NH and CoA N2'' CoA is nearly immobile along its entire length during all stages of the enzyme reaction; native and C-terminally His6-tagged wild-type enzymes in complexes including freeze-trapped acetylglutamyl anhydride and glutamyl-CoA thioester adducts, hanging drop vapor diffusion method, mixing of 0.002 ml of protein solution containing 5.6 mg/ml AarC in 45 mM potassium phosphate, pH 8.0, 90 mM potassium chloride, and 2 mM CoA or 6.0 mg/ml His6-tagged AarC in 45 mM Tris-HCl, pH 8.0, 90 mM potassium chloride, and 2 mM CoA, with 0.002 ml of reservoir solution containing 0.8-1.0 M sodium citrate, 0.1 M imidazole, pH 8.2, and 25 mM 2-mercaptoethanol for orthorhombic crystals or 1.7-2.0 M ammonium sulfate, 0.2 M sodium chloride, 0.1 M sodium cacodylate, pH 6.5, and 25 mM 2-mercaptoethanol for hexagonal crystals, room temperature of about 22°C, X-ray diffraction structure determination and analysis
Acetobacter aceti
Engineering
Amino acid exchange
Commentary
Organism
E294A
complete loss of activity; site-directed mutagenesis, the mutant specific catalytic activity is 10000fold reduced compared to the wild-type enzyme, ligand bound crystal structure modeling
Acetobacter aceti
E435A
mutant protein is completely insoluble; site-directed mutagenesis, the mutant is completely insoluble, ligand bound crystal structure determination and analysis, the mutant crystallizes in a closed complex containing dethiaacetyl-CoA, which adopts an unusual curled conformation
Acetobacter aceti
E435D
activity similar to wild-type; site-directed mutagenesis, the mutant catalytic properties are nearly equivalent to those of the His6-tagged wild-type enzyme, ligand bound crystal structure modeling
Acetobacter aceti
E435Q
mutant protein is completely insoluble; site-directed mutagenesis, the mutant is completely insoluble, ligand bound crystal structure modeling
Acetobacter aceti
N347A
large decrease in catalytic activity; site-directed mutagenesis, the mutant shows impaired catalytic activity, but the apparent affinities for all four substrates are largely unaffected, ligand bound crystal structure modeling
Acetobacter aceti
R228E
large decrease in catalytic activity; site-directed mutagenesis, the mutant has a specific defect in its ability to bind both carboxylate substrates, ligand bound crystal structure modeling
Acetobacter aceti
S71A
large decrease in catalytic activity; site-directed mutagenesis, the mutant shows impaired catalytic activity associated with lower kcat values, ligand bound crystal structure modeling
Acetobacter aceti
Inhibitors
Inhibitors
Commentary
Organism
Structure
acetate
-
Acetobacter aceti
citrate
weak competitive inhibition against succinate
Acetobacter aceti
succinate
weak competitive inhibition
Acetobacter aceti
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
steady-state kinetic analysis using a Michaelis-Menten model, a substrate inhibition model, or a competitive inhibition model, overview. Arg228 has an important kinetic role in carboxylate substrate binding
Acetobacter aceti
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
succinyl-CoA + acetate
Acetobacter aceti
via an acetylglutamyl anhydride intermediate and glutamyl-CoA thioester adduct
acetyl-CoA + succinate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Acetobacter aceti
B3EY95
; gene aarC
-
Acetobacter aceti 1023
B3EY95
gene aarC
-
Reaction
Reaction
Commentary
Organism
succinyl-CoA + acetate = acetyl-CoA + succinate
the general half-reaction for class I CoA-transferases shows two tetrahedral oxyanion intermediates, which differ by whether CoA becomes attached to the external carbonyl, provided by the acyl-CoA/carboxylate substrate, or the internal carbonyl, provided by the essential active-site glutamate. Following exchange of the carboxylate product, the second half-reaction proceeds in the reverse order of the first half-reaction. In the first half-reaction, the binary enzyme·acyl-CoA complex is converted into a CoA thiolate complex that also contains an acylglutamyl anhydride adduct. Ping-pong kinetic mechanism. Val270 has a dual influence on carboxylate substrate selectivity, as a gate and as a clamp, Arg228 has an important kinetic role in carboxylate substrate binding. The auxiliary site nonselectively binds carboxylates at the threshold of the catalytic pocket, while selectivity is enforced by the conserved gating residue Val270 and the interior of the catalytic pocke
Acetobacter aceti
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + acetoacetate
0.35% of the activity with succinate
721675
Acetobacter aceti
acetoacetyl-CoA + acetate
-
-
-
r
acetyl-CoA + D-malate
0.28% of the activity with succinate
721675
Acetobacter aceti
D-malyl-CoA + acetate
-
-
-
r
acetyl-CoA + fumarate
0.20% of the activity with succinate
721675
Acetobacter aceti
fumaryl-CoA + acetate
-
-
-
r
acetyl-CoA + propionate
0.34% of the activity with succinate
721675
Acetobacter aceti
propionyl-CoA + acetate
-
-
-
r
acetyl-CoA + succinate
-
721675
Acetobacter aceti
succinyl-CoA + acetate
-
-
-
r
additional information
no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti
?
-
-
-
-
additional information
substrate specificity, overview. Acetoacetate, propionate, D-malate, fumarate, L-malate, formate, oxaloacetate, DL-methylsuccinate, glutarate are alternate substrates, no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti
?
-
-
-
-
additional information
no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti 1023
?
-
-
-
-
additional information
substrate specificity, overview. Acetoacetate, propionate, D-malate, fumarate, L-malate, formate, oxaloacetate, DL-methylsuccinate, glutarate are alternate substrates, no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti 1023
?
-
-
-
-
succinyl-CoA + acetate
5.1% of the activity with succinate + acetyl-CoA
721675
Acetobacter aceti
acetyl-CoA + succinate
-
-
-
r
succinyl-CoA + acetate
via an acetylglutamyl anhydride intermediate and glutamyl-CoA thioester adduct
721675
Acetobacter aceti
acetyl-CoA + succinate
-
-
-
?
succinyl-CoA + acetoacetate
-
721675
Acetobacter aceti
acetoacetyl-CoA + succinate
-
-
-
?
succinyl-CoA + D-malate
-
721675
Acetobacter aceti
D-malyl-CoA + succinate
-
-
-
?
succinyl-CoA + DL-methylsuccinate
-
721675
Acetobacter aceti
DL-methylsuccinyl-CoA + succinate
-
-
-
?
succinyl-CoA + formate
-
721675
Acetobacter aceti
formyl-CoA + succinate
-
-
-
?
succinyl-CoA + fumarate
-
721675
Acetobacter aceti
fumaryl-CoA + succinate
-
-
-
?
succinyl-CoA + glutarate
-
721675
Acetobacter aceti
glutaryl-CoA + succinate
-
-
-
?
succinyl-CoA + L-malate
-
721675
Acetobacter aceti
L-malyl-CoA + succinate
-
-
-
?
succinyl-CoA + oxaloacetate
-
721675
Acetobacter aceti
oxaloacetyl-CoA + succinate
-
-
-
?
succinyl-CoA + propionate
-
721675
Acetobacter aceti
propionyl-CoA + succinate
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Acetobacter aceti
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Acetobacter aceti
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
3.8
6.8
-
Acetobacter aceti
3.8
-
unstable below
Acetobacter aceti
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
150
-
citrate
pH 8.0, temperature not specified in the publication
Acetobacter aceti
150
-
succinate
pH 8.0, 30°C
Acetobacter aceti
1600
-
acetate
pH 8.0, temperature not specified in the publication
Acetobacter aceti
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli; gene aarC, sequence comparisons and phylogenetic analysis, expression of C-terminally His6-tagged wild-type and mutant enzymes
Acetobacter aceti
Crystallization (Commentary) (protein specific)
Crystallization
Organism
crystal structures of a C-terminally His6-tagged form of several wild-type and mutant complexes, including freeze-trapped acetylglutamyl anhydride and glutamyl-CoA thioester adducts. The latter shows the acetate product bound to an auxiliary site that is required for efficient carboxylate substrate recognition. Mutant E294A crystallizes in a closed complex containing dethiaacetyl-CoA, which adopts an unusual curled conformation. A model of the acetyl-CoA Michaelis complex reveals that the nucleophilic glutamate is held at a near-ideal angle for attack as the thioester oxygen is forced into an oxyanion hole composed of Gly388 NH and CoA N2'' CoA is nearly immobile along its entire length during all stages of the enzyme reaction; native and C-terminally His6-tagged wild-type enzymes in complexes including freeze-trapped acetylglutamyl anhydride and glutamyl-CoA thioester adducts, hanging drop vapor diffusion method, mixing of 0.002 ml of protein solution containing 5.6 mg/ml AarC in 45 mM potassium phosphate, pH 8.0, 90 mM potassium chloride, and 2 mM CoA or 6.0 mg/ml His6-tagged AarC in 45 mM Tris-HCl, pH 8.0, 90 mM potassium chloride, and 2 mM CoA, with 0.002 ml of reservoir solution containing 0.8-1.0 M sodium citrate, 0.1 M imidazole, pH 8.2, and 25 mM 2-mercaptoethanol for orthorhombic crystals or 1.7-2.0 M ammonium sulfate, 0.2 M sodium chloride, 0.1 M sodium cacodylate, pH 6.5, and 25 mM 2-mercaptoethanol for hexagonal crystals, room temperature of about 22°C, X-ray diffraction structure determination and analysis
Acetobacter aceti
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E294A
complete loss of activity; site-directed mutagenesis, the mutant specific catalytic activity is 10000fold reduced compared to the wild-type enzyme, ligand bound crystal structure modeling
Acetobacter aceti
E435A
mutant protein is completely insoluble; site-directed mutagenesis, the mutant is completely insoluble, ligand bound crystal structure determination and analysis, the mutant crystallizes in a closed complex containing dethiaacetyl-CoA, which adopts an unusual curled conformation
Acetobacter aceti
E435D
activity similar to wild-type; site-directed mutagenesis, the mutant catalytic properties are nearly equivalent to those of the His6-tagged wild-type enzyme, ligand bound crystal structure modeling
Acetobacter aceti
E435Q
mutant protein is completely insoluble; site-directed mutagenesis, the mutant is completely insoluble, ligand bound crystal structure modeling
Acetobacter aceti
N347A
large decrease in catalytic activity; site-directed mutagenesis, the mutant shows impaired catalytic activity, but the apparent affinities for all four substrates are largely unaffected, ligand bound crystal structure modeling
Acetobacter aceti
R228E
large decrease in catalytic activity; site-directed mutagenesis, the mutant has a specific defect in its ability to bind both carboxylate substrates, ligand bound crystal structure modeling
Acetobacter aceti
S71A
large decrease in catalytic activity; site-directed mutagenesis, the mutant shows impaired catalytic activity associated with lower kcat values, ligand bound crystal structure modeling
Acetobacter aceti
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
acetate
-
Acetobacter aceti
citrate
weak competitive inhibition against succinate
Acetobacter aceti
succinate
weak competitive inhibition
Acetobacter aceti
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
150
-
citrate
pH 8.0, temperature not specified in the publication
Acetobacter aceti
150
-
succinate
pH 8.0, 30°C
Acetobacter aceti
1600
-
acetate
pH 8.0, temperature not specified in the publication
Acetobacter aceti
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
steady-state kinetic analysis using a Michaelis-Menten model, a substrate inhibition model, or a competitive inhibition model, overview. Arg228 has an important kinetic role in carboxylate substrate binding
Acetobacter aceti
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
succinyl-CoA + acetate
Acetobacter aceti
via an acetylglutamyl anhydride intermediate and glutamyl-CoA thioester adduct
acetyl-CoA + succinate
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + acetoacetate
0.35% of the activity with succinate
721675
Acetobacter aceti
acetoacetyl-CoA + acetate
-
-
-
r
acetyl-CoA + D-malate
0.28% of the activity with succinate
721675
Acetobacter aceti
D-malyl-CoA + acetate
-
-
-
r
acetyl-CoA + fumarate
0.20% of the activity with succinate
721675
Acetobacter aceti
fumaryl-CoA + acetate
-
-
-
r
acetyl-CoA + propionate
0.34% of the activity with succinate
721675
Acetobacter aceti
propionyl-CoA + acetate
-
-
-
r
acetyl-CoA + succinate
-
721675
Acetobacter aceti
succinyl-CoA + acetate
-
-
-
r
additional information
no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti
?
-
-
-
-
additional information
substrate specificity, overview. Acetoacetate, propionate, D-malate, fumarate, L-malate, formate, oxaloacetate, DL-methylsuccinate, glutarate are alternate substrates, no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti
?
-
-
-
-
additional information
no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti 1023
?
-
-
-
-
additional information
substrate specificity, overview. Acetoacetate, propionate, D-malate, fumarate, L-malate, formate, oxaloacetate, DL-methylsuccinate, glutarate are alternate substrates, no activity with glycolate, glyoxylate, oxalate, trifluoroacetate, DL-lactate, L-lactate, malonate, pyruvate, maleate, butyrate, D-tartrate, L-tartrate, 2-oxooglutarate, citrate, or DL-isocitrate
721675
Acetobacter aceti 1023
?
-
-
-
-
succinyl-CoA + acetate
5.1% of the activity with succinate + acetyl-CoA
721675
Acetobacter aceti
acetyl-CoA + succinate
-
-
-
r
succinyl-CoA + acetate
via an acetylglutamyl anhydride intermediate and glutamyl-CoA thioester adduct
721675
Acetobacter aceti
acetyl-CoA + succinate
-
-
-
?
succinyl-CoA + acetoacetate
-
721675
Acetobacter aceti
acetoacetyl-CoA + succinate
-
-
-
?
succinyl-CoA + D-malate
-
721675
Acetobacter aceti
D-malyl-CoA + succinate
-
-
-
?
succinyl-CoA + DL-methylsuccinate
-
721675
Acetobacter aceti
DL-methylsuccinyl-CoA + succinate
-
-
-
?
succinyl-CoA + formate
-
721675
Acetobacter aceti
formyl-CoA + succinate
-
-
-
?
succinyl-CoA + fumarate
-
721675
Acetobacter aceti
fumaryl-CoA + succinate
-
-
-
?
succinyl-CoA + glutarate
-
721675
Acetobacter aceti
glutaryl-CoA + succinate
-
-
-
?
succinyl-CoA + L-malate
-
721675
Acetobacter aceti
L-malyl-CoA + succinate
-
-
-
?
succinyl-CoA + oxaloacetate
-
721675
Acetobacter aceti
oxaloacetyl-CoA + succinate
-
-
-
?
succinyl-CoA + propionate
-
721675
Acetobacter aceti
propionyl-CoA + succinate
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Acetobacter aceti
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Acetobacter aceti
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
3.8
6.8
-
Acetobacter aceti
3.8
-
unstable below
Acetobacter aceti
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the class I-CoA-transferases, which, typified by mitochondrial succinyl-CoA:3-oxoacid CoA-transferase, form multiple covalent adducts involving an essential glutamate residue. Arg228 is found in only AarC and several closely allied SCACT group sequences, EC 6.2.1
Acetobacter aceti
additional information
the nucleophilic glutamate is held at a near-ideal angle for attack as the thioester oxygen is forced into an oxyanion hole composed of Gly388 NH and CoA N2''. CoA is nearly immobile along its entire length during all stages of the enzyme reaction. Spatial and sequence conservation of key residues indicates that this mechanism is general among class I CoA-transferases, structural model for the AarC mechanism, overview. An auxiliary carboxylate binding site, located just outside the AarC catalytic pocket, contributes to the efficient recognition and conversion of the physiological carboxylate substrates. Protein conformational dynamics, overview. Arg228 has an important kinetic role in carboxylate substrate binding. Regulation of carboxylate access to the active-site glutamate, overview
Acetobacter aceti
physiological function
the enzyme is an acetic acid resistance factor AarC that is required for acetate resistance by vinegar factory strain Acetobacter aceti 1023. The enzyme acts in a variant citric acid cycle that overoxidizes acetic acid to CO2, which then diffuses into the acidic culture medium
Acetobacter aceti
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the class I-CoA-transferases, which, typified by mitochondrial succinyl-CoA:3-oxoacid CoA-transferase, form multiple covalent adducts involving an essential glutamate residue. Arg228 is found in only AarC and several closely allied SCACT group sequences, EC 6.2.1
Acetobacter aceti
additional information
the nucleophilic glutamate is held at a near-ideal angle for attack as the thioester oxygen is forced into an oxyanion hole composed of Gly388 NH and CoA N2''. CoA is nearly immobile along its entire length during all stages of the enzyme reaction. Spatial and sequence conservation of key residues indicates that this mechanism is general among class I CoA-transferases, structural model for the AarC mechanism, overview. An auxiliary carboxylate binding site, located just outside the AarC catalytic pocket, contributes to the efficient recognition and conversion of the physiological carboxylate substrates. Protein conformational dynamics, overview. Arg228 has an important kinetic role in carboxylate substrate binding. Regulation of carboxylate access to the active-site glutamate, overview
Acetobacter aceti
physiological function
the enzyme is an acetic acid resistance factor AarC that is required for acetate resistance by vinegar factory strain Acetobacter aceti 1023. The enzyme acts in a variant citric acid cycle that overoxidizes acetic acid to CO2, which then diffuses into the acidic culture medium
Acetobacter aceti
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.000074
-
succinyl-CoA
pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.00013
-
succinyl-CoA
pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.00039
-
succinyl-CoA
pH 8.0, 30°C, mutant N347A
Acetobacter aceti
0.0007
-
acetyl-CoA
pH 8.0, 30°C, mutant N347A; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.0023
-
acetyl-CoA
pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.0034
-
acetyl-CoA
pH 8.0, 30°C, His6-tagged wild-type enzyme
Acetobacter aceti
0.005
-
acetyl-CoA
pH 8.0, 30°C, mutant E435D
Acetobacter aceti
0.009
-
succinyl-CoA
pH 8.0, 30°C, His6-tagged wild-type enzyme and mutant E435D
Acetobacter aceti
0.029
-
acetate
mutant S71A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.03
-
succinate
mutant S71A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.074
-
succinyl-CoA
mutant S71A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.13
-
succinyl-CoA
mutant R228E, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.16
-
acetate
mutant N347A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant N347A
Acetobacter aceti
0.3
-
succinate
mutant R228E, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.39
-
succinyl-CoA
mutant N347A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.6
-
acetate
mutant R228E, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.7
-
acetyl-CoA
mutant N347A, pH 8.0, temperature not specified in the publication; mutant S71A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
2.7
-
succinate
mutant N347A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant N347A
Acetobacter aceti
3
-
acetate
mutant E435D, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant E435D
Acetobacter aceti
3.4
-
acetyl-CoA
wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
4
-
acetate
pH 8.0, 30°C, His6-tagged wild-type enzyme; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
5.2
-
acetyl-CoA
mutant E435D, pH 8.0, temperature not specified in the publication
Acetobacter aceti
9
-
succinyl-CoA
mutant E435D, pH 8.0, temperature not specified in the publication; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
73
-
succinate
mutant E435D, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant E435D
Acetobacter aceti
79
-
succinate
pH 8.0, 30°C, His6-tagged wild-type enzyme; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.000074
-
succinyl-CoA
pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.00013
-
succinyl-CoA
pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.00039
-
succinyl-CoA
pH 8.0, 30°C, mutant N347A
Acetobacter aceti
0.0007
-
acetyl-CoA
pH 8.0, 30°C, mutant N347A; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.0023
-
acetyl-CoA
pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.0034
-
acetyl-CoA
pH 8.0, 30°C, His6-tagged wild-type enzyme
Acetobacter aceti
0.005
-
acetyl-CoA
pH 8.0, 30°C, mutant E435D
Acetobacter aceti
0.009
-
succinyl-CoA
pH 8.0, 30°C, His6-tagged wild-type enzyme and mutant E435D
Acetobacter aceti
0.029
-
acetate
mutant S71A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.03
-
succinate
mutant S71A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant S71A
Acetobacter aceti
0.074
-
succinyl-CoA
mutant S71A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.13
-
succinyl-CoA
mutant R228E, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.16
-
acetate
mutant N347A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant N347A
Acetobacter aceti
0.3
-
succinate
mutant R228E, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.39
-
succinyl-CoA
mutant N347A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
0.6
-
acetate
mutant R228E, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant R228E
Acetobacter aceti
0.7
-
acetyl-CoA
mutant N347A, pH 8.0, temperature not specified in the publication; mutant S71A, pH 8.0, temperature not specified in the publication
Acetobacter aceti
2.7
-
succinate
mutant N347A, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant N347A
Acetobacter aceti
3
-
acetate
mutant E435D, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant E435D
Acetobacter aceti
3.4
-
acetyl-CoA
wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
4
-
acetate
pH 8.0, 30°C, His6-tagged wild-type enzyme; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
5.2
-
acetyl-CoA
mutant E435D, pH 8.0, temperature not specified in the publication
Acetobacter aceti
9
-
succinyl-CoA
mutant E435D, pH 8.0, temperature not specified in the publication; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
73
-
succinate
mutant E435D, pH 8.0, temperature not specified in the publication; pH 8.0, 30°C, mutant E435D
Acetobacter aceti
79
-
succinate
pH 8.0, 30°C, His6-tagged wild-type enzyme; wild-type, pH 8.0, temperature not specified in the publication
Acetobacter aceti
Other publictions for EC 2.8.3.18
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)
738287
Murphy
Functional dissection of the b ...
Acetobacter aceti 1023, Acetobacter aceti
Front. Chem.
4
23
2016
-
-
1
1
1
-
-
-
-
-
-
2
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
721675
Mullins
Crystal structures of Acetobac ...
Acetobacter aceti 1023, Acetobacter aceti
Biochemistry
51
8422-8434
2012
-
-
1
1
7
-
3
1
-
-
-
1
-
4
-
-
-
1
-
-
-
-
20
-
1
-
-
-
1
-
2
-
3
-
-
-
-
1
-
1
7
-
-
3
3
1
-
-
-
1
-
-
-
-
-
-
-
-
20
-
1
-
-
-
1
-
2
-
-
3
3
-
25
25
692865
Mullins
A specialized citric acid cycl ...
Acetobacter aceti
J. Bacteriol.
190
4933-4940
2008
-
-
1
-
-
-
3
8
-
-
2
-
-
2
-
-
1
1
-
-
4
-
6
1
1
-
-
7
1
-
-
-
5
1
-
-
-
2
-
-
-
-
-
5
8
12
-
-
2
-
-
-
-
1
-
-
4
-
6
1
1
-
-
12
1
-
-
1
-
1
1
-
5
5
693049
van Grinsven
Acetate:succinate CoA-transfer ...
Trichomonas vaginalis
J. Biol. Chem.
283
1411-1418
2008
-
-
1
-
-
-
-
1
1
-
1
-
-
6
-
-
-
-
-
-
1
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
-
1
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
662286
Riviere
Acetyl:succinate CoA-transfera ...
Trypanosoma brucei
J. Biol. Chem.
279
45337-45346
2004
-
1
1
-
-
-
-
-
-
-
1
1
-
2
-
-
-
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-
1
3
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
1
3
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
723135
Steinbüchel
Anaerobic pyruvate metabolism ...
Tritrichomonas foetus, Tritrichomonas foetus ATCC 30924
Mol. Biochem. Parasitol.
20
57-65
1986
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-
-
-
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1
1
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2
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2
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1
1
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-
-
-
-
-
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2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-