Substrate binding sites of Leuconostoc dextransucrase evaluated by inhibition kinetics.

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  • KOBAYASHI Mikihiko
    Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
  • YOKOYAMA Ikuko
    Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
  • MATSUDA Kazuo
    Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

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  • Substrate binding site of Leuconostoc dextransucrase evaluated by inhibition kinetics

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Graphical analysis of inhibition kinetics for dextransucrase from Leuconostoc mesenteroides was done with typical inhibitors, competitive and noncompetitive. Based on the plots of Yonetani-Theorell and Semenza-Balthazar, mutual competition between the pairs of inhibitors of identical kinetic type was observed, while combination of competitive and noncompetitive inhibitors gave no significant mutual interactions. By the procedure of Nitta et al., binding sites for competitive and noncompetitive inhibitors were shown to be distant from each other. Moreover, two noncompetitive inhibitors competed with each other for a single binding site on the enzyme. Although biphasic reciprocal plots may suggest rather complicated binding of various inhibitors, the results obtained by the three graphical methods are fully explained when competitive and noncompetitive inhibitors for substrate sucrose bind to the so-called donor- and acceptor-sites of dextransucrase, respectively.

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