title: Pyruvate decarboxylase from Kluyveromyces lactis. An enzyme with an extraordinary substrate activation behaviour.
abstract: Pyruvate decarboxylase (EC 4.1.1.1) was isolated and purified from the yeast Kluyveromyces lactis. The properties of this enzyme relating to the native oligomeric state, the subunit size, the nucleotide sequence of the coding gene(s), the catalytic activity, and protein fluorescence as well as circular dichroism are very similar to those of the well characterized pyruvate decarboxylase species from yeast. Remarkable differences were found in the substrate activation behaviour of the two pyruvate decarboxylases using three independent methods: steady-state kinetics, stopped-flow measurements, and kinetic dilution experiments. The dependence of the observed activation rate constant on the substrate concentration of pyruvate decarboxylase from K. lactis showed a minimum at a pyruvate concentration of 1.5 mm. According to the mechanism of substrate activation suggested this local minimum occurs due to the big ratio of the dissociation constants for the binding of the first (regulatory) and the second (catalytic) substrate molecule. The microscopic rate constants of the substrate activation could be determined by a refined fit procedure. The influence of the artificial activator pyruvamide on the activation of the enzyme was studied.
authors.affiliation: Institut für Biochemie, Fachbereich Biochemie/Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06099 Halle/Saale, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Krieger
authors.forename: Michael
authors.initials: M
authors.lastname: Spinka
authors.forename: Ralph
authors.initials: R
authors.lastname: Golbik
authors.forename: Gerhard
authors.initials: G
authors.lastname: Hübner
authors.forename: Stephan
authors.initials: S
authors.lastname: König
mesh_terms: D020134:Catalytic Domain; D004789:Enzyme Activation; D007700:Kinetics; D007716:Kluyveromyces / Q000201:enzymology*; D011767:Pyruvate Decarboxylase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019289:Pyruvic Acid / Q000737:chemistry / Q000378:metabolism
chemical_list: D019289:Pyruvic Acid; D011767:Pyruvate Decarboxylase