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Literature summary for 2.3.3.3 extracted from

  • Kim, M.; Le, H.; McInerney, M.J.; Buckel, W.
    Identification and characterization of re-citrate synthase in Syntrophus aciditrophicus (2013), J. Bacteriol., 195, 1689-1696.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 CodonPlus (DE3)-GroEL cells Syntrophus aciditrophicus

Inhibitors

Inhibitors Comment Organism Structure
2-oxoglutarate 40% inhibition at 10 mM Syntrophus aciditrophicus
4-hydroxymercuribenzoate complete inhibition at 0.1 mM Syntrophus aciditrophicus
5,5'-dithiobis-2-nitrobenzoate 30% inhibition at 10 mM Syntrophus aciditrophicus
ATP 30% inhibition at 10 mM Syntrophus aciditrophicus
citrate 40% inhibition at 10 mM Syntrophus aciditrophicus
EDTA complete inhibition at 0.2 mM Syntrophus aciditrophicus
additional information not inhibited by 5 mM NADH Syntrophus aciditrophicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.085
-
oxaloacetate at pH 8.0 and 22°C Syntrophus aciditrophicus
0.13
-
acetyl-CoA at pH 8.0 and 22°C Syntrophus aciditrophicus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ the enzyme contains stoichiometric amounts of Ca2+ (0.9 Ca/73 kDa) Syntrophus aciditrophicus
Co2+ Co2+ increases the activity by about 70% compared with that obtained withMn2+ Syntrophus aciditrophicus
Mg2+ Mg2+ has no effect at all on enzyme activity Syntrophus aciditrophicus
Mn2+ assays are performed in the presence of 0.2 mM MnCl2 Syntrophus aciditrophicus
Zn2+ the EDTA-inactivated enzyme regains activity by addition Zn2+ at final concentrations of 0.2 mM Syntrophus aciditrophicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
65000
-
4 * 65000, SDS-PAGE Syntrophus aciditrophicus
71800
-
4 * 71800, calculated from amino acid sequence Syntrophus aciditrophicus
72891
-
4 * 72891, electrospray-time of flight mass spectrometry Syntrophus aciditrophicus
271000
-
gel filtration Syntrophus aciditrophicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetyl-CoA + H2O + oxaloacetate Syntrophus aciditrophicus specific substrates (R)-citrate + CoA
-
?
acetyl-CoA + H2O + oxaloacetate Syntrophus aciditrophicus ATCC 700169 specific substrates (R)-citrate + CoA
-
?

Organism

Organism UniProt Comment Textmining
Syntrophus aciditrophicus
-
-
-
Syntrophus aciditrophicus ATCC 700169
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Strep-tactin column chromatography and Superdex 200 gel filtration Syntrophus aciditrophicus

Source Tissue

Source Tissue Comment Organism Textmining
culture condition:crotonate-grown cell
-
Syntrophus aciditrophicus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.72
-
purified enzyme, in the absence of Mn2+, at pH 8.0 and 22°C Syntrophus aciditrophicus
1.2
-
purified enzyme, in the presence of 0.2 mM Mn2+, at pH 8.0 and 22°C Syntrophus aciditrophicus

Storage Stability

Storage Stability Organism
-80°C, the oxically purified enzyme, 20% loss of activity after each cycle of freezing at -80°C and thawing Syntrophus aciditrophicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + H2O + oxaloacetate specific substrates Syntrophus aciditrophicus (R)-citrate + CoA
-
?
acetyl-CoA + H2O + oxaloacetate specific substrates Syntrophus aciditrophicus ATCC 700169 (R)-citrate + CoA
-
?
additional information no activity with propionyl-CoA, 2-oxoglutarate, pyruvate, and 2-oxo-3-methylbutanoate Syntrophus aciditrophicus ?
-
?
additional information no activity with propionyl-CoA, 2-oxoglutarate, pyruvate, and 2-oxo-3-methylbutanoate Syntrophus aciditrophicus ATCC 700169 ?
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 65000, SDS-PAGE Syntrophus aciditrophicus
homotetramer 4 * 71800, calculated from amino acid sequence Syntrophus aciditrophicus
homotetramer 4 * 72891, electrospray-time of flight mass spectrometry Syntrophus aciditrophicus

Synonyms

Synonyms Comment Organism
Re-citrate synthase
-
Syntrophus aciditrophicus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
-80
-
the oxically purified enzyme loses 20% of its activity after each cycle of freezing at -80°C and thawing Syntrophus aciditrophicus