Immunological detection of AcAMP antimicrobial peptide secreted by Aspergillus clavatus

Ehsan Zamani; Jamil Zargan; Hossein Honar; Abbas Hajizade; Ashkan  Haji Noor Mohammadi; Hani  Keshavarz Alikhani; Ahmad Heidari; Mohammad  Hossein Pour

doi:10.18502/ijm.v13i2.5985

Ehsan Zamani Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran
Jamil Zargan Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran
Hossein Honar Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran
Abbas Hajizade Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran
Ashkan Haji Noor Mohammadi Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran
Hani Keshavarz Alikhani Department of Biology, Razi University, Kermanshah, Iran
Ahmad Heidari Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
Mohammad Hossein Pour Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran

DOI: https://doi.org/10.18502/ijm.v13i2.5985

Keywords: Fungi; Aspergillus; Antimicrobial peptides; Recombinant proteins; Enzyme-linked immunosorbent assay

Abstract

Background and Objectives: Aspergillus clavatus antimicrobial peptide (AcAMP) is a fungi-derived peptide with a broad spectrum of activity against pathogenic bacteria and fungi. Natural antimicrobial peptides, including AcAMP, have attracted many attentions in the development of new natural antibiotics against pathogenic bacteria, especially multidrug resistant ones.

Materials and Methods: In the present study, acamp gene was codon-optimized and chemically synthesized in pUC57 cloning vector, subcloned into pET28a (+) expression vector and transferred into competent Escherichia coli BL21 (DE3) cells. The expression of AcAMP was induced by addition of Isopropyl β- d-1-thiogalactopyranoside (IPTG) and the expressed peptide was purified by Ni-NTA. BALB/c mice were immunized with the purified peptide and the ability of the immunized mice sera for the detection of the native AcAMP secreted by A. clavatus IRAN 142C was examined through ELISA and Western blotting techniques.

Results: Both ELISA and Western blotting demonstrated the ability of the sera of the immunized mice to detect the native AcAMP.

Conclusion: The results of the present work show that the raised antibody against recombinant AcAMP can be used to detect AcAMP peptide, an issue which paves the way to develop detection kits for the detection of AcAMP-producing organisms, purification of this valuable peptide for further investigations.