Expression of enterocin E-760 in the fusion form with SUMO-3 in Pichia pastoris X33 and its antimicrobial characterization
摘要
Recently, the application of bioadditives such as bacteriocins in food processing has received significant attention to replace chemical additives and maintain a safe and sustainable food supply. Among these, enterocin E-760 is a notable bacteriocin originated Enterrococcus sp., exhibiting great potential for food preservation due to its potent activity against both Gram-negative and Gram-positive bacteria, especially food-borne pathogens. In this study, the gene encoding enterocin E-760 was codon-optimized, fused with the sequence encoding the human SUMO-3 protein (507 bp) and inserted into pPICZαA for expression in P. pastoris. Mut+ integrants were screened via PCR and peptide expression was induced using methanol. After 48 h of induction, the culture supernatant of recombinant Pichia pastoris exhibited antilisterial activity of 80 AU mL−1. The recombinant enterocin E-760 was detected via SDS-PAGE with an estimated molecular weight of approximately 10 kDa. Furthermore, this recombinant peptide exhibited significant antimicrobial activity against several foodborne strains, such as Bacillus subtilis ATCC 6633, Bacillus cereus ATCC 10876 and Staphylococcus aureus ATCC 6538, with the activity level reaching up 320 AU mL−1. In conclusion, the antimicrobial peptide enterocin E-760 fused with SUMO protein was sucessfully expressed in P. pastoris X33 and demonstrated strong antimicrobial activity against some pathogenic bacterial strains. These findings highlight its potential applications in food preservation. However, further purification and characterization are required to enhance its effectiveness, particularly against harmful Gram-negative bacteria.