Beneficial impacts of incorporating the non‐natural amino acid azulenyl‐alanine into the Trp‐rich antimicrobial peptide buCATHL4B

Antimicrobial peptides (AMPs) present a promising scaffold for the development of po-tent antimicrobial agents. Substitution of tryptophan by non‐natural amino acid Azulenyl‐Alanine (AzAla) would allow studying the mechanism of action of AMPs by using unique properties of this amino acid, such as ability to be excited separately from tryptophan in a multi‐Trp AMPs and envi-ronmental insensitivity. In this work, we investigate the effect of Trp→AzAla substitution in anti-microbial peptide buCATHL4B (contains three Trp side chains). We found that antimicrobial and bactericidal activity of the original peptide was preserved, while cytocompatibility with human cells and proteolytic stability was improved. We envision that AzAla will find applications as a tool for studies of the mechanism of action of AMPs. In addition, incorporation of this non‐natural amino acid into AMP sequences could enhance their application properties.