| Summary: | In this study, a wide pool of lactic acid bacteria strains deposited in two recognized culture collections was tested against ropy bread spoilage bacteria, specifically belonging to <i>Bacillus</i> spp., <i>Paenibacillus</i> spp., and <i>Lysinibacillus</i> spp. High-throughput and ex vivo screening assays were performed to select the best candidates. They were further investigated to detect the production of active antimicrobial metabolites and bacteriocins. Moreover, technological and safety features were assessed to value their suitability as biocontrol agents for the production of clean-label bakery products. The most prominent inhibitory activities were shown by four strains of <i>Lactiplantibacillus plantarum</i> (NFICC19, NFICC 72, NFICC163, and NFICC 293), two strains of <i>Pediococcus pentosaceus</i> (NFICC10 and NFICC341), and <i>Leuconostoc citreum</i> NFICC28. Moreover, the whole genome sequencing of the selected LAB strains and the in silico analysis showed that some of the strains contain operons for bacteriocins; however, no significant evidence was observed phenotypically.
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