Production of Recombinant Antimicrobial Polymeric Protein Beta Casein-E 50-52 and Its Antimicrobial Synergistic Effects Assessment with Thymol.
| Autor: | Fahimirad S; Agriculture and Natural Resources Biotechnology Department, University of Tehran, Karaj 31587-11167, Iran. sh_fahimirad@yahoo.com., Abtahi H; 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. abtahi@arakmu.ac.ir., Razavi SH; Agriculture and Natural Resources Biotechnology Department, University of Tehran, Karaj 31587-11167, Iran., Alizadeh H; Agriculture and Natural Resources Biotechnology Department, University of Tehran, Karaj 31587-11167, Iran., Ghorbanpour M; Department of Medicinal Plants, Faculty of Agriculture and Natural Resources, Arak University, Arak 3815688349, Iran. |
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| Jazyk: | angličtina |
| Zdroj: | Molecules (Basel, Switzerland) [Molecules] 2017 May 31; Vol. 22 (6). Date of Electronic Publication: 2017 May 31. |
| DOI: | 10.3390/molecules22060822 |
| Abstrakt: | 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. Competing Interests: The authors declare no conflict of interest. |
| Databáze: | MEDLINE |
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