One-step enzymatic synthesis of nucleosides from low water-soluble purine bases in non-conventional media

The effect of several water-miscible cosolvents on activity and stability of soluble and immobilized 2′-deoxyribosyltransferase from Lactobacillus reuteri on Sepabeads® has been studied in order to establish optimal conditions for enzymatic synthesis of nucleosides using purine bases with low solubility in aqueous buffer. As a rule of thumb, there was a general reduction of soluble enzyme activity when cosolvent content was gradually increased in reaction medium. In contrast, immobilized enzyme activity was enhanced 1.2–1.4-fold at 20% of methanol, ethanol, 2-propanol, diethylene glycol, and acetone; and at 10% and 30% acetonitrile. Likewise, highest increased activity (1.8-fold) was also obtained in presence of 20% acetonitrile. Immobilized enzyme was successfully used in the synthesis of 2′-deoxyxanthosine and 2′-deoxyguanosine using 2′-deoxyuridine as sugar donor and the corresponding poor water-soluble base in the presence of 30% of methanol, ethanol, 2-propanol, ethylene glycol, acetonitrile, and DMSO, giving high nucleoside yields at 4 h.

► Soluble 2′-deoxyribosyltransferase activity was reduced in non-conventional media.
► Activity of immobilized enzyme was enhanced (from 1.2 to 1.8-fold) in such media.
► Stabilization against enzyme denaturation was achieved by immobilization.
► High yields of 2′-deoxyxanthosine and 2′-deoxyguanosine were achieved at short times.