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

Citation
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
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology
Journal title
ARCHIVES OF MICROBIOLOGY
ISSN journal
0302-8933 → ACNP
Volume
175
Issue
4
Year of publication
2001
Pages
308 - 312
Database
ISI
SICI code
0302-8933(200104)175:4<308:SHEAPO>2.0.ZU;2-1
Abstract
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.