CLONING OF RECOMBINANT PURINE NUCLEOSIDE PHOSPHORYLASE IN E.COLI

Purine nucleoside phosphorylase is a key enzyme in the metabolism of purine nucleosides. It catalyzes the reversible phosphorolytic cleavage of glycosidic bonds in both ribonucleosides and deoxyribonucleosides, forming purine bases in combination with ribose-1-phosphate or deoxyribose-1-phosphate. The high capacity for enzymatic trans-glycosylation of PNP from E. coli is of particular interest in biotechnology for the production of biologically active nucleoside analogues. In this work, we describe the construction of a recombinant plasmid DNA - pET-28a(+)-PNP (6070 bp), containing the PNP gene (728 bp) for expression of this enzyme in E. coli.