High-Yield Soluble Expression and Simple Purification of the Antimicrobial Peptide OG2 Using the Intein System in Escherichia coli

摘要: OG2 is a modified antimicrobial peptide, that is, derived from the frog peptide Palustrin-OG1. It has high activity and low cytotoxicity, it therefore promising as therapeutic agent. Both prokaryotic (Escherichia coli) eukaryotic (Pichia pastoris) production host systems were used to produce in our previous study; however, was difficult achieve expression yields efficient purification. In this study, we achieved high-yield using intein fusion system. The optimized gene cloned into pTWIN1 vector generate pTWIN-OG2-intein2 (C-terminal vector) pTWIN-intein1-OG2 (N-terminal vector). Nearly 70% of expressed OG2-intein2 soluble after IPTG concentration induction temperature decreased, whereas only 42% intein1-OG2 soluble. Up 75 mg obtained 1 l culture, 85% protein cleaved by 100 mM DTT. Intein1-OG2 less amenable cleavage due inhibition N-terminal amino acid OG2. purified exhibited strong against E. coli K88. system best currently available for cost-effective