<i>Lactococcus lactis</i> plays a vital role as a starter culture in dairy fermentations, exhibits remarkable protein secretion capabilities, and is Generally Regarded As Safe (GRAS). A limitation for the use of this organism for industrial and probiotic applications is the shortage of inducible gene expression systems, an essential tool for heterologous protein expression. Using a native homologous recombination and a unique in trans antibiotic selection system, we have inserted several promoter-transcription factor pairs for orthogonal expression of target proteins. The scar-less integration of these inducible gene expression systems lends itself to iterative functionalization of different inducible gene expression systems. In particular enabled induced expression from three different orthogonal inducers. Furthermore, the methods used to introduce these gene expression systems uses a mechanism that which lends its use in lactic acid bacteria beyond <i>L. lactis</i>.

Genome engineering for expanded inducible gene expression in lactic acid bacteria

ACS Fall 2020 Virtual Meeting

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