Generation of an engineered food-grade Lactococcus lactis strain for production of an antimicrobial peptide: in vitro and in silico evaluation

摘要

The schematic maps of recombinant pAMJ1653 vector (R-pAMJ1653) harboring cLFchimera and schematic representation of cLFchimera and its parts. P170 promoter, promoter is up-regulated by low-pH; SP, signal sequence of SP310mut2; LFchimera, chimeric antimicrobial peptide, T, terminator; alr WCFS1, WCFS1 gene encoding alanine racemase; repB, a replicon from for maintenance in p15A, a replicon from for maintenance in Restriction mapping analysis. ) Undigested pAMJ1653 plasmid (Lane 1 and 2, refer to replicates); ) Double digestion of recombinant pAMJ1653 vector by and . The size of the band of interest after double digestion is 125 bp. SDS-PAGE analysis of the culture supernatant. A protein band with the size of 4.2 kDa (shown by arrow) represents the recombinant cLFchimera peptide. From left to right: lane 1: size marker protein (ladder protein), lane 2: purified cLFchimera from culture supernatant of harboring recombinant pAMJ1653 vector and lane 3: culture supernatant of harboring self-ligated pAMJ1653 vector with no cLFchimera coding sequence was passed through Ni-NTA agarose column and was loaded on SDS-PAGE as negative control