title: Purification and Characterization of a Cysteine-Rich 14-kDa Antibacterial Peptide from the Granular Hemocytes of Mangrove Crab Episesarma tetragonum and Its Antibiofilm Activity.
abstract: Antimicrobial peptide (AMP) crustin is a type of immune molecule present in the immune system of crustaceans and response against microbial invasion. In the present study, we have identified and characterized the cationic, amphipathic structure consisting of AMP crustin from a mangrove crab Episesarma tetragonum using CM Sepharose-based cation exchange column chromatography. E. tetragonum crustin showed a single band of 14 kDa on SDS-PAGE and the homogeneity showed retention time of 8.4 min in RP-HPLC. Functional studies of E. tetragonum crustin exhibits the antibacterial activity (2-4 μg/ml) and biofilm inhibition (20 μg/ml) against the Gram-positive bacteria Staphylococcus aureus and Enterococcus faecalis. Hydrophobicity and extrapolysaccharide production of Gram-positive bacteria were inhibited through the bactericidal inhibitory concentration. In situ visualization analysis of biofilm inhibition was observed through light and confocal laser scanning microscopy. Surface morphology and the bacterial biofilm inhibition were viewed by scanning electron and atomic force microscopy. This study emphasizes the potential activity of E. tetragonum crustin, an interesting candidate for the development of novel broad-spectrum antimicrobial agent against bacterial pathogens. Graphical Abstract Antimicrobial peptide synthesis and host-pathogen interaction lead to production of immune molecules directed to destruction of pathogens.
authors.affiliation: Crustacean Molecular Biology & Genomics Lab, Department of Animal Health and Management, Alagappa University, Karaikudi, 630 004, Tamil Nadu, India.
authors.forename: Jeyachandran
authors.initials: J
authors.lastname: Sivakamavalli
authors.forename: Raju
authors.initials: R
authors.lastname: Nirosha
authors.forename: Baskaralingam
authors.initials: B
authors.lastname: Vaseeharan
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D003386:Brachyura / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D013293:Enterococcus faecalis / Q000187:drug effects* / Q000254:growth & development; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects; D006434:Hemocytes / Q000737:chemistry*; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008826:Microbial Sensitivity Tests; D008970:Molecular Weight; D011135:Polysaccharides, Bacterial / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D017433:Protein Structure, Secondary; D051381:Rats; D012756:Sheep; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000254:growth & development
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D011135:Polysaccharides, Bacterial