Journal
JOURNAL OF GENETIC ENGINEERING AND BIOTECHNOLOGY
Volume 21, Issue 1, Pages -Publisher
SPRINGERNATURE
DOI: 10.1186/s43141-023-00478-w
Keywords
Anti-lipopolysaccharide factor; Portunus pelagicus; Crustacean immunity; Cytotoxicity; Antimicrobial peptides
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A novel ALF protein (Pp-ALF) was identified and characterized from the blue swimmer crab, which demonstrated potential antibacterial, antiviral, antifungal, anticancer, and antibiofilm activities. However, the toxic effect of Pp-ALF on host cells prevented successful expression in Escherichia coli. Fluorescent staining showed that Pp-ALF induced host cell death by altering cell membrane permeability.
BackgroundInvertebrates like crabs employ their own immune systems to fight against a number of invasive infections. Anti-lipopolysaccharide factors (ALFs) are an important class of antimicrobial peptides (AMPs) exhibiting binding and neutralizing activities against lipopolysaccharides.ResultsThis study identified and characterized a novel homolog of ALF (Pp-ALF) from the blue swimmer crab Portunus pelagicus. Pp-ALF has a 369bp open-reading frame encoding a protein with 123 amino acids. The deduced protein featured an LPS-binding domain and a signal peptide. The predicted tertiary structure of Pp-ALF contains three alpha helices packed against four beta sheets. The deduced amino acid sequence of Pp-ALF had a net positive charge of +10.75 and an isoelectric point of 9.8. Phylogenetic analysis revealed that Pp-ALF has a strong ancestral relationship with crab ALFs.ConclusionAntibacterial, antiviral, antifungal, anticancer, and antibiofilm activities of Pp-ALF could be revealed by in silico prediction tools. Recombinant expression of Pp-ALF was unsuccessful in the Escherichia coli Rosetta-gami expression system due to the cytotoxic effect of the peptide to the host. The toxic effect of Pp-ALF to the host was displayed by membrane permeabilization and death of the host cells by fluorescent staining with Syto9-Propidium Iodide and CTC-DAPI- FITC.
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