A Novel Antimicrobial Peptide Spampcin56–86 from Scylla paramamosain Exerting Rapid Bactericidal and Anti-Biofilm Activity In Vitro and Anti-Infection In Vivo

• Published in: International journal of molecular sciences 2022-11, Vol.23 (21), p.13316
• Main Authors: Jiang, Manyu, Chen, Roushi, Zhang, Jingrong, Chen, Fangyi, Wang, Ke-Jian
• Format: Article
• Language: English
• Subjects: Amino acids; Animals; anti-biofilm; anti-infection in vivo; Antibacterial activity; Antibacterial agents; Antibiotics; Antimicrobial activity; Antimicrobial agents; antimicrobial peptide; Antimicrobial peptides; Aquaculture; Bacteria; Bacterial infections; Bactericidal activity; Biocompatibility; Biofilms; Bioinformatics; Cell death; Crustaceans; Cytotoxicity; Danio rerio; Drug resistance; E coli; Gene expression; Gram-negative bacteria; Gram-positive bacteria; In vivo methods and tests; Investigations; Marine animals; Microorganisms; mud crab Scylla paramamosain; Pathogens; Peptides; Pseudomonas aeruginosa; Sapmpcin; Scanning electron microscopy; Scylla; Scylla paramamosain; Staphylococcus infections; Thermal stability; Zebrafish
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms232113316

The abuse of antibiotics leads to the increase of bacterial resistance, which seriously threatens human health. Therefore, there is an urgent need to find effective alternatives to antibiotics, and antimicrobial peptides (AMPs) are the most promising antibacterial agents and have received extensive attention. In this study, a novel potential AMP was identified from the marine invertebrate Scylla paramamosain and named Spampcin. After bioinformatics analysis and AMP database prediction, four truncated peptides (Spa31, Spa22, Spa20 and Spa14) derived from Spampcin were screened, all of which showed potent antimicrobial activity with different antibacterial spectrum. Among them, Spampcin56–86 (Spa31 for short) exhibited strong bactericidal activity against a variety of clinical pathogens and could rapidly kill the tested bacteria within minutes. Further analysis of the antibacterial mechanism revealed that Spa31 disrupted the integrity of the bacterial membrane (as confirmed by scanning electron microscopy observation, NPN, and PI staining assays), leading to bacterial rupture, leakage of cellular contents (such as elevated extracellular ATP), increased ROS production, and ultimately cell death. Furthermore, Spa31 was found to interact with LPS and effectively inhibit bacterial biofilms. The antibacterial activity of Spa31 had good thermal stability, certain ion tolerance, and no obvious cytotoxicity. It is worth noting that Spa31 could significantly improve the survival rate of zebrafish Danio rerio infected with Pseudomonas aeruginosa, indicating that Spa31 played an important role in anti-infection in vivo. This study will enrich the database of marine animal AMPs and provide theoretical reference and scientific basis for the application of marine AMPs in medical fields.