Purification and properties of citrate lyase from Streptococcus diacetilactis.
Citrate lyase from Streptococcus diacetilactis has been purified to yield a protein that was homogeneous as judged by sedimentation velocity and sedimentation equilibrium experiments. The enzyme's sedimentation coefficient is 16.8 S and its molecular weight is around 585,000. It contains three nonidentical subunits of about 53,000, 34,000, and 10,000 daltons. The enzyme in its active form contains an acetyl group which turns over during the citrate cleavage reaction. Removal of the acetyl group inactivates the enzyme. The deacetyl enzyme can be partially reactivated by acetylation with acetic anhydride. The enzyme undergoes slow "reaction-inactivation." The rate of inactivation is first order and the rate constant of inactivation is much lower than that for a similar inactivation process of the citrate lyase from Klebsiella aerogenes. Like the latter enzyme it contains stoichiometric amounts of phosphopantothenate. The enzyme is inactivated at pH greater than 8.1 and the presence of citrate provides protection against this inactivation. Sedimentation studies of the enzyme at pH 8.7 indicate that the enzyme is dissociated, which may account for the inactivation. The enzyme is immunologically different from citrate lyases of K. aerogenes and Escherichia coli.
D001275:ATP Citrate (pro-S)-Lyase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000085:Acetates; D000818:Animals; D002850:Chromatography, Gel; D002951:Citrates / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007700:Kinetics; D007709:Klebsiella / Q000201:enzymology; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D007652:Oxo-Acid-Lyases / Q000302:isolation & purification*; D011485:Protein Binding; D011489:Protein Denaturation; D011817:Rabbits / Q000276:immunology; D013045:Species Specificity; D013291:Streptococcus / Q000201:enzymology*; D013997:Time Factors; D015032:Zinc / Q000494:pharmacology
D000085:Acetates; D002951:Citrates; D046911:Macromolecular Substances; D001275:ATP Citrate (pro-S)-Lyase; D007652:Oxo-Acid-Lyases; D008274:Magnesium; D015032:Zinc