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Isolation and Purification of an Antibacterial Protein From Immune Induced Haemolymph of American Cockroach, Periplaneta Americana



Basseri HR1 ; Dadikhoeni A2 ; Bakhtiari R3 ; Abolhassani M1 ; Hajihosseinibaghdadabadi R2
Authors
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Authors Affiliations
  1. 1. Department of Medical Entomology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Biology, Payame Noor University, Tehran, Iran
  3. 3. Department of Photobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Source: Journal of Arthropod-Borne Diseases Published:2016

Abstract

Background: Antimicrobial peptides play a role as effectors substances in the immunity of vertebrate and invertebrate hosts. In the current study, antimicrobial peptide was isolated from the haemolymph of the American cockroach, Periplaneta americana. Methods: Micrococcus luteus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria were candidate for injection. Induction was done by injecting both bacteria into the abdominal cavity of two groups of cockroaches separately. The haemolymphs were collected 24 hours after post injection and initially tested against both bacteria. Subsequently, the immune induced haemolymph was purified by high performance liquid chromatography (HPLC) to separate the proteins responsible for the antibacterial activity. Results: The non-induced haemolymph did not show any activity against both bacteria whereas induced haemolymph exhibited high activity against M. luteus but did less against E. coli. Two fractions showed antibacterial activity against M. luteus. Finally the molecular weight of the isolated antibacterial proteins were determined as 72 kDa and 62 kDa using SDS-PAGE. Conclusion: Induced haemolymph of American cockroaches has the ability to produce peptides to combat against Gram-positive bacteria when an immune challenge is mounted. Further work has to be done to sequence of the protein, which it would be advantageous.