Large quantities of antimicrobial peptides are required for investigations and clinical trials, therefore suitable production method alternative to traditional chemical synthesis is necessary. Production of recombinant antimicrobial peptides in prokaryotic systems has successfully demonstrated the viability of this approach. Production of antimicrobial peptides in Escherichia coli is potentially limited due to their toxicity to host cells and susceptibility to proteolytic degradation, which can be avoided using fusion protein approach. We describe antimicrobial peptide production in E. coli based on forcing antimicrobial peptides into inclusion bodies, which is affective for the production of large quantities of antimicrobial peptides. Chemical reagents for cleaving peptide bond between antimicrobial peptides and fusion proteins such as cyanogen bromide and diluted acid are selective and provide antimicrobial peptides for biological studies in short time.