Cost-Effective Expression and Purification of Recombinant Venom Peptide Mastoparan B of Vespa Basalis in Escherichia Coli

Abstract

Mastoparan B is a cationic, amphiphilic tetradecapeptide toxin isolated from the venom of the black-bellied hornet (Vespa basalis), the most dangerous species of wasps found in Taiwan. Mastoparan B was evaluate possess antibacterial activity and cardiovascular depress activity. However, mastoparan B is low abundance in the venom (3.4% of crude venom). In this study, mastoparan B was overexpressed in Escherichia coli BL21 as a recombinant protein fused the C-terminus of oleosin by a linker polypeptide, intein S. Artifical oil bodies (AOBs) were reconstituted with triacylglycerol, phospholipid to obtain the insoluble recombinant protein. Mastoparan B was subsequently released through self-splicing of intein induced by temperature alteration from artifical oli bodies, and the recombinant mastoparan B was collected it in the supernatant after centrifugation. Recombinant mastoparan B release from AOBs was exhibited membrane permeabilization activity, bacteriostatic and bactericidal activity. These results have shown that mastoparan B was successfully expressed and purified via the efficient AOB expression/purification system.