In this paper, lipase was used as the imprinting object. The lipase was firstly bio-imprinted, then the lipase of the imprinted enzyme was acetylated and cross-linked, and the acylation and cross-linking were detected by trinitrobenzenesulfonic acid (TNBS). degree. The results showed that the number of free amino groups decreased after acylation and cross-linking, indicating that some amino groups were involved in acetylation, which made the cross-linking of the imprinted enzyme possible. At the same time, the cross-linked enzyme was structurally stable, not easy to be inactivated, and the activity was significantly higher than that of acetylase.