Pervaporation dehydration was applied to the lipase-catalyzed esterification of fructose/glucose with palmitic acid in 2-methyl-2-butanol with the intention of improving the reaction yield. The water content in the reaction mixture decreased to 0.03 wt% by means of pervaporation using a zeolite NaA membrane. With the aid of pervaporation dehydration, the total yield of fructose monopalmitate and dipalmitate reached 66% in the esterification started with 54 mM fructose and 108 mM palmitic acid. It was possible to decrease the ratio of diester in the reaction product by decreasing the ratio of fatty acid to sugar in the substrate solution.