R. Ronimus et al., Sequencing, high-level expression and phylogeny of the pyrophosphate-dependent phosphofructokinase from the thermophilic spirochete Spirochaeta thermophila, ARCH MICROB, 175(4), 2001, pp. 308-312
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.