title: A novel single-domain peptide, anti-LPS factor from prawn: synthesis of peptide, antimicrobial properties and complete molecular characterization.
abstract: In this study, we reported a complete molecular characterization including bioinformatics features, gene expression, peptide synthesis and its antimicrobial activities of an anti-lipopolysaccharide (LPS) factor (ALF) cDNA identified from the established cDNA library of freshwater prawn Macrobrachium rosenbergii (named as MrALF). The mature protein has an estimated molecular weight of 11.240 kDa with an isoelectric point of 9.46. The bioinformatics analysis showed that the MrALF contains an antimicrobial peptide (AMP) region between T54 and P77 with two conserved cysteine residues (Cys55 and Cys76) which have an anti-parallel β-sheet confirmation. The β-sheet is predicted as cationic with hydrophobic nature containing a net charge of +5. The depicted AMP region is determined to be amphipathic with a predicted hydrophobic face 'FPVFI'. A highest MrALF gene expression was observed in hemocytes and is up-regulated with virus [white spot syndrome baculovirus (WSBV)], bacteria (Aeromonas hydrophila) and Escherichia coli LPS at various time points. The LPS binding region of MrALF peptide was synthesized to study the antimicrobial property, bactericidal efficiency and hemolytic capacity. The peptide showed antimicrobial activity against both the Gram-negative and Gram-positive bacteria. The bactericidal assay showed that the peptide recognized the LPS of bacterial cell walls and binding on its substrate and thereby efficiently distinguishing the pathogens. The hemolytic activity of MrALF peptide is functioning in a concentration dependant manner. In summary, the comprehensive analysis of MrALF showed it to be an effective antimicrobial peptide and thus it plays a crucial role in the defense mechanism of M. rosenbergii.
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India. Electronic address: jesuaraj@hotmail.com.
authors.forename: Jesu
authors.initials: J
authors.lastname: Arockiaraj
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India.
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Kumaresan
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India.
authors.forename: Prasanth
authors.initials: P
authors.lastname: Bhatt
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Palanisamy
authors.affiliation: Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX 78712, USA.
authors.forename: Annie J
authors.initials: AJ
authors.lastname: Gnanam
authors.affiliation: Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226 031, Uttar Pradesh, India.
authors.forename: Mukesh
authors.initials: M
authors.lastname: Pasupuleti
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Semeling, Bedong, Kedah 08100, Malaysia.
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Kasi
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur 603 203, Chennai, Tamil Nadu, India.
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Chaurasia
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000494:pharmacology*; D006461:Hemolysis / Q000187:drug effects; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors*; D051379:Mice; D008969:Molecular Sequence Data; D033541:Palaemonidae / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid
chemical_list: D000890:Anti-Infective Agents; D008070:Lipopolysaccharides; D010455:Peptides
keywords: Anti-lipopolysaccharide; Antimicrobial peptide; Hemolysis; Pathogen; Prawn