Factors affecting the activity of citrate synthase of Acetobacter xylinum and its possible regulatory role.
The citrate synthase activity of Acetobacter xylinum cells grown on glucose was the same as of cells grown on intermediates of the tricarboxylic acid cycle. The activity of citrate synthase in extracts is compatible with the overall rate of acetate oxidation in vivo. The enzyme was purified 47-fold from sonic extracts and its molecular weight was determined to be 280000 by gel filtration. It has an optimum activity at pH 8.4. Reaction rates with the purified enzyme were hyperbolic functions of both acetyl-CoA and oxaloacetate. The Km for acetyl-CoA is 18 mum and that for oxaloacetate 8.7 mum. The enzyme is inhibited by ATP according to classical kinetic patterns. This inhibition is competitive with respect to acetyl-CoA (Ki = 0.9 mM) and non-competitive with respect to oxaloacetate. It is not affected by changes in pH and ionic strength and is not relieved by an excess of Mg2+ ions. Unlike other Gram-negative bacteria, the A. xylinum enzyme is not inhibited by NADH, but is inhibited by high concentrations of NADPH. The activity of the enzyme varies with energy charge in a manner consistent with its role in energy metabolism. It is suggested that the flux through the tricarboxylic acid cycle in A. xylinum is regulated by modulation of citrate synthase activity in response to the energy state of the cells.
D000105:Acetyl Coenzyme A; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002950:Citrate (si)-Synthase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D003065:Coenzyme A / Q000494:pharmacology; D041822:Gluconacetobacter xylinus / Q000201:enzymology*; D005947:Glucose; D006863:Hydrogen-Ion Concentration; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D010071:Oxaloacetates; D007652:Oxo-Acid-Lyases / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D013386:Succinates
D006902:Hydroxymercuribenzoates; D010071:Oxaloacetates; D000644:Quaternary Ammonium Compounds; D013386:Succinates; D009243:NAD; D009249:NADP; D000105:Acetyl Coenzyme A; D000255:Adenosine Triphosphate; D002950:Citrate (si)-Synthase; D007652:Oxo-Acid-Lyases; D008274:Magnesium; D005947:Glucose; D003065:Coenzyme A