Lipase-catalysed alcoholysis and hydrolysis of 2＇,3＇,5＇-tri-O-acetyluridine (1a) and 2＇,3＇,5＇-tri-O-acetyl-2＇-C-methyluridine (1b) were studied. Conditions for full and regioselective deacetylation of 1a and 1b are shown in the present work. New compound 2＇,3＇-di-O-acetyl-2＇-C-methyluridine (3b) was prepared by regioselective lipase-catalysed deacetylation.