This study investigates the use of lipases for the synthesis of ester prodrugs from racemic 9-(2,3-dihydroxypropyl)adenine (DHPA) in dimethylformamide. Lipases from Geotrichum candidum 4013 were immobilized on chitosan beads and tested alongside commercially available lipases (from hog pancreas, Aspergillus niger, Candida antarctica, Pseudomonas fluorescens, and Mucor miehei). The most efficient enzyme for the reaction was Candida antarctica lipase, which achieved the highest chemical yield. However, immobilized lipases from G. candidum 4013 showed lower activity than their free forms. The study also evaluated the reusability of immobilized lipases, finding that they retained 70% of their initial activity after two cycles. The results suggest that immobilization can improve enzyme stability and reusability, which is important for industrial applications. The study highlights the potential of lipases in the synthesis of prodrugs, particularly for the production of ester prodrugs from DHPA. The findings indicate that the choice of lipase and immobilization method significantly affects the efficiency and yield of the reaction. The study also emphasizes the importance of optimizing reaction conditions to achieve high chemical yields and enantioselectivity. Overall, the research demonstrates the effectiveness of lipases in the synthesis of ester prodrugs and provides insights into the factors that influence their performance.This study investigates the use of lipases for the synthesis of ester prodrugs from racemic 9-(2,3-dihydroxypropyl)adenine (DHPA) in dimethylformamide. Lipases from Geotrichum candidum 4013 were immobilized on chitosan beads and tested alongside commercially available lipases (from hog pancreas, Aspergillus niger, Candida antarctica, Pseudomonas fluorescens, and Mucor miehei). The most efficient enzyme for the reaction was Candida antarctica lipase, which achieved the highest chemical yield. However, immobilized lipases from G. candidum 4013 showed lower activity than their free forms. The study also evaluated the reusability of immobilized lipases, finding that they retained 70% of their initial activity after two cycles. The results suggest that immobilization can improve enzyme stability and reusability, which is important for industrial applications. The study highlights the potential of lipases in the synthesis of prodrugs, particularly for the production of ester prodrugs from DHPA. The findings indicate that the choice of lipase and immobilization method significantly affects the efficiency and yield of the reaction. The study also emphasizes the importance of optimizing reaction conditions to achieve high chemical yields and enantioselectivity. Overall, the research demonstrates the effectiveness of lipases in the synthesis of ester prodrugs and provides insights into the factors that influence their performance.