Synthesis of 2'-deoxynucleosides by transglycosylation with new immobilized and stabilized uridine phosphorylase and purine nucleoside phosphorylase

Multimeric uridine phosphorylase (UP) and purine nucleoside phosphorylase (PNP) of Bacillus subtilis have been expressed from genes cloned in Escherichia coli, purified, characterized, immobilized and stabilized on solid support. A new immobilization strategy has been developed for UP onto Sepabeads coated with polyethyleneamine followed by crosslinking with aldehyde-dextran. PNP has been immobilized onto glyoxyl-agarose. At pH 10 and 45 °C these derivatives catalyzed the transglycosylation of 2’-deoxyuridine to 2’-deoxyguanosine in high yield (92%). Under the same conditions the not immobilized enzymes were promptly inactivated.