Cloning and expression of gene encoding enterocin E-760 in Escherichia coli Rosetta 2
Keywords:
Bacteriocin, Enterocin E-760, Enterococcus sp, pET22b( ), E. coli Rosetta 2Abstract
Enterocin E-760 is an antimicrobial peptide belonging to bacteriocins class II which shows a broad spectrum against both gram-negative and gram-positive especially food pathogens such as Listeria monocytogenes, Salmonella, Staphylococcus aureus, etc. This article illustrated the expression of the recombinant enterocin E-760 in E. coli Rosetta 2 with high antibacterial activity. The recombinant strain was induced in LB medium shaking at 200 rpm/min and 37 ºC and using 0.1 mM IPTG as inducer for 5 hours. The molecular weight of enterocin E-760 conjugated protein SUMO was 25 kDa by using SDS-PAGE electrophoresis of gel 12,6%; the antimicrobial activity against L. monocytogenes ATCC® 35152™ at the concentration of 105 CFU/mL reached 106 AU/mL by analyzing on agar diffusion. These results are the first step to optimize the expression process to produce a large amount of enterocin E-760 from E. coli with the purpose of applying in food preservation.
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