Phospholipase A2 (PLA2) hydrolyzes phospholipids at sn-2 position to release free fatty acids. PLA2 consists of a superfamily mainly categorized as secretory PLA2 (sPLA2), cellular Ca 2+ -dependent PLA2 (cPLA2), and cellular Ca 2+ -independent PLA2 (iPLA2). We are the first to report iPLA2 in insect. Here an objective of our study is to purify a recombinant iPLA2 protein (SeiPLA2) of Spodoptera exigua using bacterial expression system. An open reading frame of SeiPLA2 was cloned into an expression vector and then transformed into Escherichia coli BL21. Over-expression with IPTG yielded recombinant SeiPLA2 (rSeiPLA2), which was then purified by an affinity chromatography using Ni-NTA column. The purified rSeiPLA2 gave significant PLA2 activity using a pyrene substrate. Its activity was inhibited by an iPLA2 specific inhibitor (BEL), but not by sPLA2 inhibitor (BPB) or cPLA2 inhibitor (MAFP