De-novo design and production of antimicrobial peptides

Antimicrobial peptides are important defense compounds of higher organisms that can be used as therapeutic agents against bacterial and/or viral infections. We designed peptides and demonstrated their high antimicrobial activity and low host cell cytotoxicity. With the increasing demand for antimicrobial compounds for production of “healthy” food, these peptides might serve as templates for novel antimicrobial agents. The plant protecting ability of the designed peptides was demonstrated by spraying them on the plant surface. Besides the generation of transgenic plants this way of application displays a promising method for practical use of antimicrobial peptides in plant protection.

For practical application high amounts of peptide are required. We developed a protocol for plant-based production and purification of biologically active, hydrophobic and positively charged antimicrobial peptide. We generated a tobacco mosaic virus-based transient production system following the “full virus vector strategy” to produce the antimicrobial peptide as fusion protein with the viral coat protein. After successful production in tobacco plants the modified virus was extracted from the plants and the peptide was released from the coat protein and purified. Removal of plant, viral and bacterial contaminants by high resolution purification methods enables recovery of pure and biologically active AMPs from transfected plant materials that can be used in many applications, including human and animal medicine or agriculture.

Christian Lindermayr

General workflow for production and purification of recombinant proteins/peptides in plants.