Investigating the production of antimicrobial peptides using the SUMO protein expression system in E. coli




Mersereau, Mary deLissa

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In an age of increasing antibiotic resistance, antimicrobial peptides (AMPs) are a promising alternative to antibiotics. While numerous publications have documented AMP expression, few have used the same production system to express a variety of different AMPs. In this study, eight AMPs were expressed in Escherichia coli using the SUMO carrier protein. Six of the eight AMPs were designed to target Helicobacter pylori, a gram-negative bacterium known for causing chronic gastritis and gastric cancer. The goal was to identify a universal AMP production system, assess the capabilities of the SUMO carrier protein, and identify AMPs likely to eliminate H. pylori. The two metrics used to assess expression success were yield and activity against the target bacteria. The remaining two AMPs targeted a gram-positive bacterium, Staphylococcus aureus, and showed excellent yield and activity. However, yield was poor for all six AMPs targeting H. pylori. Five of these had enough yield to conduct activity tests, but activity was low for all. Thus, we demonstrated that SUMO is an unreliable carrier protein for AMPs toxic to gram-negative bacteria. These results led the Kearney Lab to identify a superior production method, the Inclusion Body Method. Five of the AMPs targeting H. pylori were transferred to inclusion body vectors and have now been produced in good quantity. AMPs remain a promising and exciting alternative treatment to antibiotics and further exploration is needed.



AMP production system, Inclusion body method