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Effects of pH and divalent cation concentration on M. tuberculosis IspD activity. (A) The optimal pH for catalytic activity was determined using MES (pH 5.5 to 6.5), MOPS (pH 6.5 to 7.5), Tris-HCl (pH 7.5 to 8.5), and TAPS (pH 8.5 to 9.5). (B) Divalent cations (Mg2+, Mn2+, Zn2+, or Ca2+) were added to the reaction mixtures at the indicated concentrations. Reaction mixtures containing 38.5 pmol of purified IspD enzyme were prepared as described in Materials and Methods.
