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.
Synthesis of 2'-deoxynucleosides by transglycosylation with new immobilized and stabilized uridine phosphorylase and purine nucleoside phosphorylase
UBIALI, DANIELA;SCARAMOZZINO, FRANCESCA;TERRENI, MARCO;ALBERTINI, ALESSANDRA;PREGNOLATO, MASSIMO
2004-01-01
Abstract
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.File in questo prodotto:
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