A fusion protein designed for soluble expression, rapid purification, and enhanced stability of parasporin-2 with potential therapeutic applications

Abstract

Bacillus thuringiensis parasporin-2 (PS2Aa1 or Mpp46Aa1) selectively destroys human cancer cells, making it a promising anticancer agent. PS2Aa1 protoxin expression in Escherichia coli typically results in inclusion bodies that must be solubilized and digested by proteinase K to become active. Here, maltose-binding protein (MBP) was fused to the N-terminus of PS2Aa1, either full-length (MBP-fPS2) or truncated (MBP-tPS2), to increase soluble protein expression in E. coli and avoid solubilization and proteolytic activation. Soluble MBP-fPS2 and MBD-tPS2 proteins were produced in E. coli and purified with endotoxin levels below 1 EU/μg. MBP-fPS2 was cytotoxic against T cell leukemia MOLT-4 and Jurkat cell lines after proteinase-K digestion. However, MBP-tPS2 was cytotoxic immediately without MBP tag removal or activation. MBP-tPS2′s thermal stability also makes it appropriate for bioproduction and therapeutic applications.