2-Fluorocordycepin (2-fluoro-3’-deoxyadenosine) is known as anticancer and antiviral agent.New method of the chemoenzymatic synthesis of 2-fluorocordicepin (5) is suggested. Thelithium salt of 1-phosphate 3-deoxyribose (3-dRib-1-P) (4) and 2-fluoroadenosine were usedas a donor of 3-deoxy-D-ribofuranose and of 2-fluoroadenine correspondingly. The reactionwas catalyzed by recombinant E. coli purine nucleoside phosphorylase (PNP).D-Xylofuranose (1) was used as a starting compound for synthesis of 4 (Scheme). First, 1-and 2-OH groups were protected by isopropylidene residue. Subsequent acylation withthiocarbonyldiimidazole in presence of dimethylaminopyridine followed by deoxygenationleaded to protected carbohydrates 2 and 3. After full deprotection a mixture of 3-dRib-1-Pand 3-dRib-2-P of compound 4 as lithium salt was obtained. All intermediates were isolatedand characterized by mass- and NMR-spectroscopy.Compound 4 was found to be a good substrate for recombinant E. coli PNP. The optimalconditions for synthesis of the target product 5 were determined: ratio of lithium salt of 1-Р-3-dRib:2FAdo – 3:1, PNP 1050 u.a., 50 oC, 10 mM KH2PO4 (pH 8.0).The reaction time was 168 h with the maximum conversion of 2-fluoroadenosine to theproduct 57%. The semi-preparative synthesis of 2-fluorocordycepin under those conditionswas performed with 38% yield.