Enzymatic synthesis of 2′-deoxyadenosine and 6-methylpurine-2′-deoxyriboside by Escherichia coli DH5α overexpressing nucleoside phosphorylases from Escherichia coli BL21
2010
Abstract Genes encoding purine nucleoside phosphorylase ( deo D ), uridine phosphorylase ( udp ) and thimidine phosphorylase ( deo A ) from Escherichia coli BL21 were cloned and overexpressed in E. coli DH5α. The recombinant strains were employed to synthesize 2′-deoxyadenosine (dAR) and 6-methylpurine-2′-deoxyriboside (MePdR). Experimental parameters such as strains, temperature, pH, reagent concentration and cell mass were optimized. Under the optimal situation, 96% adenine was converted to dAR and 95% 6-methylpurine (MeP) was converted to MePdR in an hour, using 0.2‰ (dry wt./v) cell paste as biocatalyst.
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