Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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The pH Studies of Recombinant Acetohydroxy Acid Synthase from Tobacco

  • Choi, Jung-Do (Department of Chemistry, Hanyang University) ;
  • Kim, Bok-Hwan (School of Life Sciences, Chungbuk National University) ;
  • Yoon, Moon-Young (Department of Health Science, Korea National Open University)
  • Published : 2003.05.20
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Abstract

The pH dependence of the kinetic parameters of recombinant acetohydroxy acid synthase catalyzed reaction was determined in order to obtain information about the chemical mechanism, particularly acid-base chemistry. The maximum velocity and V/K for pyruvate were bell-shaped with estimated pK values of 6.5-6.7 and 8.6-8.9, respectively. The maximum velocity and V/K for 2-ketobutyrate were also bell-shaped with estimated pK values of 6.6-7.0 and 8.4-8.6. The pH dependence of 1/Ki for 3-bromopyruvate, a competitive inhibitor of pyruvate, was also bell-shaped, giving pK values almost identical with those obtained for pyruvate. Since the same pK values were observed in the $pK_{i 3-bromopyruvate}$, V/K pH profiles and $V_{max}$ profiles, both enzyme groups must be in their optimum protonation state for efficient binding of reactants. These results reflect that two enzyme groups are necessary for binding of substrate and/or catalysis.

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