title: ATP-independent DNA topoisomerase from Fervidobacterium islandicum.
abstract: Thermotogales are thermophilic eubacteria belonging to a very slowly evolving branch in the eubacterial tree. In this report, we describe the purification and characterization of an ATP-independent DNA topoisomerase from the Thermotogale, Fervidobacterium islandicum. The enzyme, a monomer of about 75 kDa, is a type I DNA topoisomerase sharing many properties with the other bacterial topoisomerases I: it absolutely requires Mg2+ for activity, relaxes negatively but not positively supercoiled DNA and is inhibited by single-stranded M13 DNA and spermidine. A feature of the F. islandicum ATP-independent DNA topoisomerase I is its thermophily. The optimal temperature for the enzymatic activity is 75 degrees C. Studies about thermostability show that the enzyme is more stable when incubated undiluted in the storage buffer. In these conditions, 60% activity was retained after a 30 min preincubation at 75 degrees C.
authors.affiliation: Laboratoire d'Enzymologie des Acides Nucléiques, URA 1354 Centre National de la Recherche Scientifique, Université Paris-Sud, Orsay, France.
authors.forename: C
authors.initials: C
authors.lastname: Bouthier de la Tour
authors.forename: C
authors.initials: C
authors.lastname: Portemer
authors.forename: P
authors.initials: P
authors.lastname: Forterre
authors.forename: R
authors.initials: R
authors.lastname: Huber
authors.forename: M
authors.initials: M
authors.lastname: Duguet
mesh_terms: D004264:DNA Topoisomerases, Type I / Q000302:isolation & purification*; D004278:DNA, Superhelical; D004795:Enzyme Stability; D008274:Magnesium; D013696:Temperature; D013822:Thermoplasma / Q000201:enzymology; D059004:Topoisomerase I Inhibitors
chemical_list: D004278:DNA, Superhelical; D059004:Topoisomerase I Inhibitors; D004264:DNA Topoisomerases, Type I; D008274:Magnesium