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Molecules 2017, 22(6), 822; doi:10.3390/molecules22060822

Production of Recombinant Antimicrobial Polymeric Protein Beta Casein-E 50-52 and Its Antimicrobial Synergistic Effects Assessment with Thymol

Agriculture and Natural Resources Biotechnology Department, University of Tehran, Karaj 31587-11167, Iran
Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak 38181-76941, Iran
Department of Medicinal Plants, Faculty of Agriculture and Natural Resources, Arak University, Arak 3815688349, Iran
Author to whom correspondence should be addressed.
Academic Editor: Philippe Bulet
Received: 24 March 2017 / Revised: 8 May 2017 / Accepted: 11 May 2017 / Published: 31 May 2017
(This article belongs to the Special Issue Bioactive Natural Peptides As A Pipeline For Therapeutics)
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Accelerating emergence of antimicrobial resistance among food pathogens and consumers’ increasing demands for preservative-free foods are two contemporary challenging aspects within the food industry. Antimicrobial packaging and the use of natural preservatives are promising solutions. In the present study, we used beta-casein—one of the primary self-assembly proteins in milk with a high polymeric film production capability—as a fusion partner for the recombinant expression of E 50-52 antimicrobial peptide in Escherichia coli. The pET21a-BCN-E 50-52 construct was transformed to E. coli BL21 (DE3), and protein expression was induced under optimized conditions. Purified protein obtained from nickel affinity chromatography was refolded under optimized dialysis circumstances and concentrated to 1600 µg/mL fusion protein by ultrafiltration. Antimicrobial activities of recombinant BCN-E 50-52 performed against Escherichia coli, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus aureus, Aspergillus flavus, and Candida albicans. Subsequently, the synergistic effects of BCN-E 50-52 and thymol were assayed. Results of checkerboard tests showed strong synergistic activity between two compounds. Time–kill and growth kinetic studies indicated a sharp reduction of cell viability during the first period of exposure, and SEM (scanning electron microscope) results validated the severe destructive effects of BCN E 50-52 and thymol in combination on bacterial cells. View Full-Text
Keywords: antimicrobial activity; beta-casein; E 50-52; synergism; thymol antimicrobial activity; beta-casein; E 50-52; synergism; thymol

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Fahimirad, S.; Abtahi, H.; Razavi, S.H.; Alizadeh, H.; Ghorbanpour, M. Production of Recombinant Antimicrobial Polymeric Protein Beta Casein-E 50-52 and Its Antimicrobial Synergistic Effects Assessment with Thymol. Molecules 2017, 22, 822.

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