Nucleoside phosphorylases (NPs; E.C. 2.4.2) catalyze the reversible cleavage of the glycosidic bond of (deoxy) ribonucleosides in the presence of inorganic orthophosphate to generate the nucleobase and a-D-(deoxy)ribose-1-phosphate. If a second nucleobase is added to the reaction medium the formation of a new nucleoside can result. We have recently reported on the production and characterization of a purine nucleoside phosphorylase from A. hydrophila (AhPNPII). This PNP has been used in the synthesis of a few 6-substituted-purine-9-ribosides. We have now isolated, cloned and expressed four new NPs from C. koseri, C. perfringens and S. pyogenes (CkPNPI, CkPNPII, CpUP and SpUP). Their substrate specificity has been investigated and compared to that of AhPNPII and other enzymes previously reported. CpUP, AhPNPII and CkPNPI were immobilized and used in the synthesis of antiviral araA and ddI.
A collection of immobilized and stabilized nucleoside phosphorylases for the synthesis of nucleoside analogues
SERRA, IMMACOLATA;UBIALI, DANIELA;TERRENI, MARCO;ALBERTINI, ALESSANDRA;
2012-01-01
Abstract
Nucleoside phosphorylases (NPs; E.C. 2.4.2) catalyze the reversible cleavage of the glycosidic bond of (deoxy) ribonucleosides in the presence of inorganic orthophosphate to generate the nucleobase and a-D-(deoxy)ribose-1-phosphate. If a second nucleobase is added to the reaction medium the formation of a new nucleoside can result. We have recently reported on the production and characterization of a purine nucleoside phosphorylase from A. hydrophila (AhPNPII). This PNP has been used in the synthesis of a few 6-substituted-purine-9-ribosides. We have now isolated, cloned and expressed four new NPs from C. koseri, C. perfringens and S. pyogenes (CkPNPI, CkPNPII, CpUP and SpUP). Their substrate specificity has been investigated and compared to that of AhPNPII and other enzymes previously reported. CpUP, AhPNPII and CkPNPI were immobilized and used in the synthesis of antiviral araA and ddI.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
