Microscopic and metabolomics analysis of the anti-Listeria activity of natural and engineered cruzioseptins

• Published in: Biochimie 2024-10, Vol.225, p.168-175
• Main Authors: Bermúdez-Puga, Sebastián, Dias, Meriellen, Lima Reis, Iara, Freire de Oliveira, Taciana, Yokomizo de Almeida, Sonia Regina, Mendes, Maria Anita, Moore, Simon J., Almeida, José R., Proaño-Bolaños, Carolina, Pinheiro de Souza Oliveira, Ricardo
• Format: Article
• Language: English
• Subjects: Antibacterial; Listeria monocytogenes; Membrane lysis; Metabolomics; Skin secretion-derived peptides
• ISSN: 0300-9084; 1638-6183; 1638-6183
• DOI: 10.1016/j.biochi.2024.05.022

Listeria monocytogenes is a human opportunistic foodborne pathogen that produces life-threatening infections with a high mortality rate. The control of Listeria in the food production environment and effective clinical management of human listeriosis are challenging due to the emergence of antibiotic resistance. Hence we evaluate the in vitro anti-Listeria activity of two synthetic cruzioseptins reproducing their natural sequences CZS-9, and CZS-12, and one engineered sequence based on CZS-1, named [K4K15]CZS-1. The assessment of the in vitro potential of cruzioseptins, highlighted the promising antibacterial effect of [K4K15]CZS-1 in very low concentrations (0.91 μM) and its thermal stability at high-temperature conditions, is compatible with the food industry. Microscopic and metabolomic analyses suggest cruzioseptin induces anti-Listeria bioactivity through membrane disruption and changes in the intracellular metabolome. We also report that [K4K15]CZS-1 is not resistant to peptidases/proteases emphasizing a key advantage for their use as a food preservative. However, there is a need for further structural and functional optimisations for the potential clinical application as an antibiotic. In conclusion, [K4K15]CZS-1 stand out as membrane-active peptides with the ability to induce shifts in the bacteria metabolome and inspire the development of strategies for the prevention of L. monocytogenes emergence and dissemination. •Frog Antimicrobial Peptides (AMPs) are an alternative bioactive for controlling listeriosis.•The cationic net charge is important to the potency of anti-Listeria activity.•Cruzioseptins can induce intracellular changes in the biological pathways of bacteria.•Engineered cruzioseptin is thermostable and retains bioactivity.