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

Abstract

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.

Anti-microbial activity and immunomodulation of recombinant hepcidin 2 and NK-lysin from flounder (Paralichthys olivaceus)

“…Many AMPs have been demonstrated to directly target and disrupt bacterial membranes. Spampcin from Scylla paramamosain permeabilize S. aureus and P. aeruginosa outer and inner membranes [45]; A type I interferon (named gcIFN-20) from grass carp was shown to adsorb to the surface of S. aureus, and had a disrupting effect on the integrity of the bacterial cell membrane [28]. In the current study, localization of rPoHep2 and rPoNKL in bacteria were determined by indirect immunofluorescence.…”

A Comprehensive Review of Recent Research into the Effects of Antimicrobial Peptides on Biofilms—January 2020 to September 2023

“…However, the publication did not include in vitro or in vivo data [131]. Finally, Jiang et al used bioinformatics to generate four truncated AMPs of Spampcin (Spa31, Spa22, Spa20, and Spa14) [132]. Spa31 demonstrated potent antimicrobial activity against E. coli, P. aeruginosa, and S. aureus, inhibited/eradicated P. aeruginosa biofilms, and significantly improved the survival rate (from 50% to 79%) of Danio rerio zebrafish infected with P. aeruginosa [132].…”

Newly Discovered Antimicrobial Peptide Scyampcin 44–63 from Scylla paramamosain Exhibits a Multitargeted Candidacidal Mechanism In Vitro and Is Effective in a Murine Model of Vaginal Candidiasis

“…Since 2006, our group has identified various novel AMPs in S. paramamosain , such as Scygonadin ( 7 ), Scyreprocin ( 8 ), Sparamosin 26–54 ( 9 ), Sparanegtin ( 10 ). and Spampcin 56–86 ( 11 ). In the present study, we identified a novel AMP named Scyampcin 44–63 from the mud crab S. paramamosain .…”