title: Macrobrachium rosenbergii mannose binding lectin: synthesis of MrMBL-N20 and MrMBL-C16 peptides and their antimicrobial characterization, bioinformatics and relative gene expression analysis.
abstract: Mannose-binding lectin (MBL), an antimicrobial protein, is an important component of innate immune system which recognizes repetitive sugar groups on the surface of bacteria and viruses leading to activation of the complement system. In this study, we reported a complete molecular characterization of cDNA encoded for MBL from freshwater prawn Macrobrachium rosenbergii (Mr). Two short peptides (MrMBL-N20: (20)AWNTYDYMKREHSLVKPYQG(39) and MrMBL-C16: (307)GGLFYVKHKEQQRKRF(322)) were synthesized from the MrMBL polypeptide. The purity of the MrMBL-N20 (89%) and MrMBL-C16 (93%) peptides were confirmed by MS analysis (MALDI-ToF). The purified peptides were used for further antimicrobial characterization including minimum inhibitory concentration (MIC) assay, kinetics of bactericidal efficiency and analysis of hemolytic capacity. The peptides exhibited antimicrobial activity towards all the Gram-negative bacteria taken for analysis, whereas they showed the activity towards only a few selected Gram-positive bacteria. MrMBL-C16 peptides produced the highest inhibition towards both the Gram-negative and Gram-positive bacteria compared to the MrMBL-N20. Both peptides do not produce any inhibition against Bacillus sps. The kinetics of bactericidal efficiency showed that the peptides drastically reduced the number of surviving bacterial colonies after 24 h incubation. The results of hemolytic activity showed that both peptides produced strong activity at higher concentration. However, MrMBL-C16 peptide produced the highest activity compared to the MrMBL-N20 peptide. Overall, the results indicated that the peptides can be used as bactericidal agents. The MrMBL protein sequence was characterized using various bioinformatics tools including phylogenetic analysis and structure prediction. We also reported the MrMBL gene expression pattern upon viral and bacterial infection in M. rosenbergii gills. It could be concluded that the prawn MBL may be one of the important molecule which is involved in antimicrobial mechanism. Moreover, MrMBL derived MrMBL-N20 and MrMBL-C16 peptides are important antimicrobial peptides for the recognition and eradication of viral and bacterial pathogens.
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: Mukesh Kumar
authors.initials: MK
authors.lastname: Chaurasia
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: Rajesh
authors.initials: R
authors.lastname: Palanisamy
authors.affiliation: Department of Zoology, Pachaiyappa's College for Men, 631 501 Kanchipuram, Tamil Nadu, India.
authors.forename: Ramasamy
authors.initials: R
authors.lastname: Harikrishnan
authors.affiliation: Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, 226031 Lucknow, 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, 08100 Bedong, Kedah, Malaysia.
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Kasi
mesh_terms: D016980:Aeromonas hydrophila / Q000187:drug effects / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000998:Antiviral Agents / Q000494:pharmacology*; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D030381:Nodaviridae / Q000502:physiology; D033541:Palaemonidae / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology / Q000821:virology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014733:Vibrio / Q000187:drug effects / Q000502:physiology; D046848:White spot syndrome virus 1 / Q000502:physiology
chemical_list: D000900:Anti-Bacterial Agents; D000998:Antiviral Agents; D060829:Arthropod Proteins; D037241:Mannose-Binding Lectins; D012333:RNA, Messenger
keywords: Bactericidal effect; Hemolysis; Lectin; Peptide; Prawn