Cloning of purine nucleoside phosphorylase II gene from Bacillus stearothermophilus TH 6-2 and characterization of its gene product.
Bacillus stearothermophilus TH 6-2 has two kinds of purine nucleoside phosphorylases (Pu-NPases). The type I enzyme (Pu-NPase I) is a functional and structural homolog of eukaryotic purine nucleoside phosphorylases that catalyze the phosphorolysis of inosine, guanosine, and their derivatives. The type II enzyme (Pu-NPase II) is a minor enzyme that efficiently phosphorolyzes adenosine and its derivatives rather than other purine nucleosides like Escherichia coli Pu-NPase. The gene coding for Pu-NPase II (punB gene) has been cloned and sequenced from TH 6-2 strain. The deduced amino acid sequence of Pu-NPase II shared 54% identity with that of E. coli enzyme, while it had no significant homology to that of Pu-NPase I or eukaryotic enzymes. By producing the Pu-NPase II in E. coli cells, the Pu-NPase II has been purified and characterized.
Hamamoto
T
T
Research & Development Division, Yamasa Corporation, Chiba, Japan.
Noguchi
T
T
Midorikawa
Y
Y
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011683:Purine-Nucleoside Phosphorylase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013696:Temperature
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D011683:Purine-Nucleoside Phosphorylase