Accéder au contenu
Merck

Molecular cloning and expression of the pyrimidine nucleoside phosphorylase gene from Bacillus stearothermophilus TH 6-2.

Bioscience, biotechnology, and biochemistry (1996-10-01)
K Okuyama, T Hamamoto, T Noguchi, Y Midorikawa
RÉSUMÉ

The pyrimidine nucleoside phosphorylase (Py-NPase) of Bacillus stearothermophilus TH 6-2 is a dimer of 46-kDa subunits and catalyzes the reversible phosphorolysis of uridine and thymidine. The gene encoding this pyrimidine nucleoside phosphorylase (pyn gene) has been cloned and sequenced from B. stearothermophilus TH 6-2. The pyn gene corresponded to an open reading frame of 1299 nucleotides that translates into a putative 433 amino acid protein with a molecular weight of 46,271. The deduced amino terminal sequence of Py-NPase coincided with that previously found for the purified enzyme. The deduced amino acid sequence of Py-NPase shared significant similarity with those of human and Escherichia coli thymidine phosphorylases. The cloned pyn gene was overexpressed in E. coli cells to produce an active enzyme in large quantities that accounted for approximately 20% of the total protein.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Pyrimidine Nucleoside Phosphorylase from Bacillus subtilis, recombinant, expressed in E. coli, ≥70 units/mg protein
Sigma-Aldrich
Pyrimidine nucleoside phosphorylase, recombinant, expressed in E. coli, ≥1300 U/mL