Open AccessPurification and properties of citrate synthase from Acetobacter europaeus
Author(s) -
Sievers Martin,
Stöckli Michèle,
Teuber Michael
Publication year - 1997
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1997.tb10170.x
Subject(s) - citrate synthase , biochemistry , enzyme , atp synthase , citric acid cycle , atp citrate lyase , biology , molecular mass , acetobacter , enzyme assay , chemistry , fermentation
Citrate synthase (EC 4.1.3.7) was purified from the acidophilic bacterium Acetobacter europaeus to electrophoretic homogeneity. The specific activity was 228 units/mg of protein during the exponential ethanol‐oxidation growth phase. The enzyme has a molecular mass of 280 kDa and is a hexamer with a subunit size of 46 kDa. The apparent K m values were 20 μM for oxaloacetate and 51 μM for acetyl‐CoA. Unlike citrate synthase from other Gram‐negative bacteria, the activity of the enzyme was inhibited by ATP, slightly enhanced by ADP and not effected by NADH. Acetate caused activation of the enzyme. The pH optimum on the citrate synthase activity in vitro was 8.1. The amino‐terminal amino acid sequence of the purified enzyme was ENGKSATISLNGKDVALPVL.