Abstract
The α-acetolactate synthase from Leuconostoc mesenteroides subsp. cremoris was purified to homogeneity in SDS-PAGE. The enzyme is a trimer of 3×55,000 Da. It was unstable but could be preserved by addition of pyruvate and thiamine pyrophosphate in the buffer. The enzyme exhibits Michaelis-Menten kinetics, and K m for pyruvate is 10 mM. Three intermediates in glucose metabolism (ATP, 3-phosphoglycerate, and phosphoenolpyruvate) exhibit a noncompetitive inhibition towards the enzyme. This enzyme does not require any divalent metal ion for activity. The α-acetolactate synthase from Leuconostoc mesenteroides subsp. cremoris is not inhibited by the branched-chain amino acids (valine, leucine, and isoleucine), is FAD independent, and displays an optimal activity at pH 5.3. Therefore, it can be concluded that the purified enzyme belongs to the catabolic α-acetolactate synthases, involved in the 2,3-butanediol pathway but not in branchedchain amino acids biosynthesis.
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Phalip, V., Schmitt, P. & Diviès, C. Purification and characterization of the catabolic α-acetolactate synthase from Leuconostoc mesenteroides subsp. cremoris . Current Microbiology 31, 316–321 (1995). https://doi.org/10.1007/BF00314587
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DOI: https://doi.org/10.1007/BF00314587