Sequencing, high-level expression and phylogeny of the pyrophosphate-dependent phosphofructokinase from the thermophilic spirochete Spirochaeta thermophila

The full-length ene encoding a 554-amino-acid, active pyrophosphate-depende nt phosphofructokinase from Spirochaeta thermophila was cloned and sequence d using a combination of degenerate and inverse PCR, and the enzyme express ed to a high level in Escherichia coli. The recombinant enzyme, with a calc ulated molecular mass of 61 kDa, was purified to near homogeneity and found to be similar to the purified native enzyme for most properties examined. Phylogenetic analysis demonstrated a close relationship between the thermop hilic S. thermophila phosphofructokinase and the large beta -subunits of th e phosphofructokinases from Borrelia burgdorferi and Treponema pallidum.