Lysophospholipase (LysoPLA) is one of the most important deacylating phospholipases. The LysoPLA gene from Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae) (ApLysoPLA), a well-known wild silkmoth, was isolated and characterized. The isolated ApLysoPLA cDNA sequence was 1151 bp, with an open reading frame of 663 bp encoding a polypeptide of 220 amino acids. The deduced ApLysoPLA protein sequence has 89 and 82% identity with Heliconius reato (L.) LysoPLA and Bombyx mori L. LysoPLA, respectively; however, it shows 66, 62, 50, and 47% identity with Tribolium castaneum (Herbst) LysoPLA, Drosophila melanogaster (Meigen) LysoPLA, Homo sapiens LysoPLA I, and Mus musculus LysoPLA I, respectively. Phylogenetic analysis indicated that lepidopteran LysoPLAs, including ApLysoPLA, might be a new member of the LysoPLA family. Semiquantitative reverse transcription-polymerase chain reaction analysis showed that the ApLysoPLA gene was transcribed during four developmental stages (egg, larva, pupa, and adult) and that it was present in all tissues tested (blood, midgut, silk glands, Malpighian tubules, spermaries, ovaries, brain, muscle, fat body, and body wall), with the most abundance in Malpighian tubules. Consequently, we refer that ApLysoPLA plays an important role in the development of A. pernyi.