Biochemical characterization of an active pyrophosphate-dependent phosphofructokinase from Treponema pallidum

Abstract

An active pyrophosphate-dependent phosphofructokinase containing a six residue polyhistidine tag has been cloned from Treponema pallidum, and characterized biochemically. The phosphofructokinase has pH optima for activity of 8.0 for both the forward and reverse reactions. The apparent Km for pyrophosphate was 0.042 mM (Vmax of 141 U mg−1 protein) and for fructose-6-phosphate, 0.529 mM. The apparent Km for the reverse reaction for fructose-1,6-diphosphate was 0.267 mM (Vmax of 42.4 U mg−1 protein). The enzyme appears to be both a dimer and non-allosteric.