Abstract: D-tagatose 3-epimerase (DTE) was immobilized on 14 kinds of ion-exchange and adsorption resins, in which D301-Ⅲ was the best material for the immobilization.Then the enzyme was immobilized by cross-linkage of glutaraldehyde.The optimum conditions for the immobilization were as follows:2.00mL/g (liquid enzyme/wet resin) , pH7.5, immobilization temperature 30℃, and immobilization time 8h.The concentration of the crosslinking agent (glutaraldehyde) was 0.05%.The crosslinking time was 3h.The activity recovery yield of immobilized enzyme was above 50%.The remaining activity was more than 65% of the primary enzyme activity after the immobilized enzyme was used repeatedly for 10 times.It showed that the immobilized DTE had good operational stability.