The juvenile hormone (JH) epoxide hydrolase (JHEH) catalyzes the degradation of JH, which regulates the metamorphosis development of insects. Here we report the 2.30 Å crystal structure of JHEH from the silkworm Bombyx mori (BmJHEH). The overall structure of BmJHEH is composed of an N-terminal segment followed by a core hydrolase domain, which is interrupted by an all-α lid domain. Structural analyses together with molecular simulation reveal insights into the conservation and specificity of the active-site pocket. These findings increase our understanding of the substrate recognition and catalysis of microsomal epoxide hydrolase family and might help the design of JH-derived pesticides. © Proteins 2014;. © 2014 Wiley Periodicals, Inc.