Enantiopure epoxides are remarkably versatile intermediates for the synthesis of numerous biologically active targets, to which considerable efforts have been devoted either chemically or biologically during the past few decades. This review will emphasize the application of biocatalysis as an efficient alternative that complements conventional chemical reactions, with a special focus on the epoxidation reactions catalyzed with monooxygenases and chloroperoxidases and the hydrolytic kinetic resolution catalyzed with epoxide hydrolases. Their scopes and limitations will be elaborately discussed as compared with their chemical counterparts. These biocatalytic approaches have not only provided environmentally friendly alternatives, but also displayed advantages for certain types of enantiopure epoxides, and could serve as potential tools for synthetic chemists.