J Pharm Pharmacogn Res 2(Suppl. 1): S7, 2014
Special supplement with the abstract book of LATINFARMA 2013
CO 012: PANUSIN, A NEW DEFENSIN-LIKE ANTIMICROBIAL PEPTIDE FROM THE HEMOCYTES OF SPINY LOBSTER Panulirus argus
Pacios-Michelena A1, Montero-Alejo V1, Perdomo-Morales R1, Corzo G2, Pardo-Ruiz Z1, Vega-Hurtado Y1
Introduction: Naturally occurring antimicrobial peptides are component of first line of host defense against pathogens as part of the humoral innate immune response in crustacean. Marine invertebrates lack an acquired immune system, and must rely mainly on innate immunity mechanisms encompassed by cellular and humoral responses. In the spiny lobsters Panulirus argus, there are yet any reports concerning the isolation and characterization of these biomolecules from hemolymph.
Material and methods: Hemocytes were analyzed after in vivo challenged of juvenile lobsters with LPS. The TFA acid precipitation was used as fractionation procedure and the acid lysate extract showing antimicrobial activity was further purified by cation exchange chromatography. Purity was evaluated using RP-HPLC and TricineUrea-PAGE electrophoresis. The minimal inhibitory concentration (MIC) was evaluated by broth microdilution assay against Gram positive and Gram negative microorganisms, while the susceptibility assay was assessed by radial diffusion. Complete amino acid sequence was established by a combination of Edman degradation, mass spectrometry and transcriptome analysis.
Results: the sequence analysis demonstrated for the first time a defensin-like antimicrobial peptide isolated from crustacean. This defensin shows potent antimicrobial activity against gram-positive and gram-negative bacteria, it is not haemolytic or LPS-binding activity. Besides, it enhances LPS-induced pro-inflammatory cytokines IL-6, IL1β and TNFα as shows in the Monocyte Activation Test (MAT).
Conclusion: A defensin-like antimicrobial peptide was identified with potent antimicrobial activity.