Discovery of a high-performance phage-derived promoter/repressor system for probiotic lactobacillus engineering

Abstract

ABSTRACTBackgroundTheLactobacillusfamily comprises many species of great importance for the food and healthcare industries, with numerous strains identified as beneficial for humans and used as probiotics. Hence, there is a growing interest in engineering these probiotic bacteria as live biotherapeutics for animals and humans. However, the genetic parts needed to regulate gene expression in these bacteria remain limited compared to model bacteria likeE. coliorB. subtilis. To address this deficit, in this study, we selected and tested several bacteriophage-derived genetic parts with the potential to regulate transcription in lactobacilli.ResultsWe screened genetic parts from 6 different lactobacilli-infecting phages and identified one promoter/repressor system with unprecedented functionality inL. plantarumWCFS1. The phage-derived promoter was found to achieve expression levels nearly 9-fold higher than the previously reported strongest promoter in this strain and the repressor was able to almost completely repress this expression by reducing it nearly 500-fold.ConclusionsThe new parts and insights gained from their engineering will enhance the genetic programmability of lactobacilli for healthcare and industrial applications.