title: Sequencing, high-level expression and phylogeny of the pyrophosphate-dependent phosphofructokinase from the thermophilic spirochete Spirochaeta thermophila.
abstract: The full-length gene encoding a 554-amino-acid, active pyrophosphate-dependent phosphofructokinase from Spirochaeta thermophila was cloned and sequenced using a combination of degenerate and inverse PCR, and the enzyme expressed to a high level in Escherichia coli. The recombinant enzyme, with a calculated 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 thermophilic S. thermophila phosphofructokinase and the large beta-subunits of the phosphofructokinases from Borrelia burgdorferi and Treponema pallidum.
authors.affiliation: Department of Biological Sciences, University of Waikato, Hamilton, New Zealand. ronimus@waikato.ac.zn
authors.forename: R
authors.initials: R
authors.lastname: Ronimus
authors.forename: E
authors.initials: E
authors.lastname: de Heus
authors.forename: A
authors.initials: A
authors.lastname: Rückert
authors.forename: H
authors.initials: H
authors.lastname: Morgan
mesh_terms: D000595:Amino Acid Sequence; D001898:Borrelia / Q000201:enzymology; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D013142:Spirochaeta / Q000201:enzymology*; D014208:Treponema / Q000201:enzymology
chemical_list: D010770:Phosphotransferases; C022502:pyrophosphate-fructose 6-phosphate 1-phosphotransferase