The scheme of enzymatic preparation of fludarabine-5′-monophosphate from 2-fluoroadenine and cyclocytidine as a donor of arabinose fragment was developed and experimentally proved. Acetylphosphate was used as a donor of phosphate group. Cytidine deaminase, uridine phosphorylase, purine nucleoside phosphorylase and deoxynucleoside kinase recovered from earlier designed strains of Escherichia coli were used as biocatalysts. The synthesized fludarabine-5′-monophosphate is planned for using as a pro-drug in enzymatic pro-medicinal approach to therapy of cancer.