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.

Immune indices and identical functions of two prophenoloxidases from the haemolymph of green tiger shrimp Penaeus semisulcatus and its antibiofilm activity

“…Haemocyanin exhibited affinity towards bacteria resulting in elimination of the microbial pathogen directly from the internal environment of the potential crustacean host. Our results get supported by Sivakamavalli et al (2015) reporting that the haemocyanin affinity towards bacteria was noticed in M. japonicus haemocyanin, which was able to agglutinate both Gram-positive and Gram-negative bacteria. Due to more prevalence on antibiotic resistance in Gram-negative bacteria, the Pp-Hc antibiofilm activity was checked against them, i.e., V. alginolyticus, V. harveyi, V. parahaemolyticus, P. aeruginosa and P. vulgaris.…”

Anti-biofilm properties and immunological response of an immune molecule lectin isolated from shrimp Metapenaeus monoceros

“…MmLec therefore has a great potential to inhibit and accumulate undesirable pathogenic products by preventing the proliferation and colonization as well as interrupting their biofilm architectural cascade. Jayanthi et al [50], Sivakamavalli et al [71] and Anjugam et al [72] have also reported the similar activities of various lectins purified from other shrimp species. They also postulated that lectins, showing antibiofilm properties can be treated as a potent antimicrobial therapeutic tool.…”

Peptide Dendrimer-Based Antibacterial Agents: Synthesis and Applications

“… 71 Specially, crustin from crustaceans and plant HDPs are rich in cysteines ( Table 1 ). 111 , 112 These amino acids and features mentioned above are often used while designing AMPs.…”