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Literature summary for 2.3.3.16 extracted from

  • Johnson, D.E.; Hanson, R.S.
    Bacterial citrate synthases: purification, molecular weight and kinetic mechanism (1974), Biochim. Biophys. Acta, 350, 336-353.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
alpha-ketoglutarate
-
Acinetobacter calcoaceticus subsp. anitratus
AMP slight activation Acinetobacter calcoaceticus subsp. anitratus

General Stability

General Stability Organism
very labile, glycerol, 20% v/v or more stabilizes Bacillus subtilis

Inhibitors

Inhibitors Comment Organism Structure
acetyl-CoA no inhibition Acinetobacter calcoaceticus subsp. anitratus
acetyl-CoA no inhibition Azotobacter vinelandii
acetyl-CoA substrate inhibition at high concentration Bacillus subtilis
ATP possibly involved in enzyme regulation Acinetobacter calcoaceticus subsp. anitratus
ATP possibly involved in enzyme regulation Azotobacter vinelandii
ATP possibly involved in enzyme regulation Bacillus subtilis
citrate competitive with acetyl-CoA, noncompetitive with oxaloacetate Acinetobacter calcoaceticus subsp. anitratus
citrate competitive with acetyl-CoA, noncompetitive with oxaloacetate Azotobacter vinelandii
citrate competitive with acetyl-CoA, noncompetitive with oxaloacetate Bacillus subtilis
CoA competitive with acetyl-CoA, noncompetitive with oxaloacetate Acinetobacter calcoaceticus subsp. anitratus
CoA competitive with acetyl-CoA, noncompetitive with oxaloacetate Azotobacter vinelandii
CoA competitive with acetyl-CoA, noncompetitive with oxaloacetate Bacillus subtilis
NADH 12% inhibition at 1 mM Acinetobacter calcoaceticus subsp. anitratus
NADH
-
Azotobacter vinelandii
NADPH 29% inhibition at 0.25 mM Acinetobacter calcoaceticus subsp. anitratus
oxaloacetate no inhibition Acinetobacter calcoaceticus subsp. anitratus
oxaloacetate no inhibition Azotobacter vinelandii
oxaloacetate substrate inhibition Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
oxaloacetate
-
Azotobacter vinelandii
0.08
-
acetyl-CoA
-
Azotobacter vinelandii
0.1
-
acetyl-CoA
-
Acinetobacter calcoaceticus subsp. anitratus
0.6
-
acetyl-CoA
-
Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
KCl
-
Acinetobacter calcoaceticus subsp. anitratus
KCl
-
Azotobacter vinelandii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
58500
-
4 * 58500, sedimentation equilibrium in guanidine-HCl and dithiothreitol Azotobacter vinelandii
60000 95000 gel filtration, sedimentation equilibrium centrifugation Bacillus subtilis
69000
-
4 * 69000, sedimentation equilibrium in guanidine-HCl and dithiothreitol Acinetobacter calcoaceticus subsp. anitratus
242000
-
sedimentation equilibrium centrifugation Acinetobacter calcoaceticus subsp. anitratus
250000
-
sedimentation equilibrium centrifugation Azotobacter vinelandii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetyl-CoA + oxaloacetate + H2O Bacillus subtilis entry step to tricarboxylic acid cycle citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O Acinetobacter calcoaceticus subsp. anitratus entry step to tricarboxylic acid cycle citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O Azotobacter vinelandii entry step to tricarboxylic acid cycle citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O Bacillus subtilis HS2A2 entry step to tricarboxylic acid cycle citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O Azotobacter vinelandii OP entry step to tricarboxylic acid cycle citrate + CoA
-
?

Organism

Organism UniProt Comment Textmining
Acinetobacter calcoaceticus subsp. anitratus
-
-
-
Azotobacter vinelandii
-
-
-
Azotobacter vinelandii OP
-
-
-
Bacillus subtilis
-
-
-
Bacillus subtilis HS2A2
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Acinetobacter calcoaceticus subsp. anitratus
-
Azotobacter vinelandii
partial Bacillus subtilis

Reaction

Reaction Comment Organism Reaction ID
acetyl-CoA + H2O + oxaloacetate = citrate + CoA random sequential reaction order mechanism Bacillus subtilis
acetyl-CoA + H2O + oxaloacetate = citrate + CoA random sequential reaction order mechanism Acinetobacter calcoaceticus subsp. anitratus
acetyl-CoA + H2O + oxaloacetate = citrate + CoA random sequential reaction order mechanism Azotobacter vinelandii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.6
-
partially purified enzyme Bacillus subtilis
52.3
-
purified enzyme Azotobacter vinelandii
235
-
purified enzyme Acinetobacter calcoaceticus subsp. anitratus
523
-
-
Azotobacter vinelandii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + oxaloacetate + H2O
-
Bacillus subtilis citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O
-
Acinetobacter calcoaceticus subsp. anitratus citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O
-
Azotobacter vinelandii citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O entry step to tricarboxylic acid cycle Bacillus subtilis citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O entry step to tricarboxylic acid cycle Acinetobacter calcoaceticus subsp. anitratus citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O entry step to tricarboxylic acid cycle Azotobacter vinelandii citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O
-
Bacillus subtilis HS2A2 citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O entry step to tricarboxylic acid cycle Bacillus subtilis HS2A2 citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O
-
Azotobacter vinelandii OP citrate + CoA
-
?
acetyl-CoA + oxaloacetate + H2O entry step to tricarboxylic acid cycle Azotobacter vinelandii OP citrate + CoA
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 58500, sedimentation equilibrium in guanidine-HCl and dithiothreitol Azotobacter vinelandii
tetramer 4 * 69000, sedimentation equilibrium in guanidine-HCl and dithiothreitol Acinetobacter calcoaceticus subsp. anitratus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
enzyme assay at Bacillus subtilis
30
-
enzyme assay at Acinetobacter calcoaceticus subsp. anitratus
30
-
enzyme assay at Azotobacter vinelandii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
942
-
citrate
-
Acinetobacter calcoaceticus subsp. anitratus
2100
-
citrate
-
Azotobacter vinelandii

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.75
-
ATP versus acetyl-CoA Azotobacter vinelandii
1
-
ATP
-
Bacillus subtilis