Caister Academic Press

Recombinant Gene Expression in Lactobacilli: Strategies and Applications

Clemens Peterbauer, Stefan Heinl, Aleš Berlec and Reingard Grabherr
from: Lactobacillus Genomics and Metabolic Engineering (Edited by: Sandra M. Ruzal). Caister Academic Press, U.K. (2019) Pages: 169-186.

Abstract

Bacteria-based expression systems play a substantial role in industrial and medical biotechnology for the production of various proteins. In contrast to Gram negative bacteria such as Escherichia coli, Gram positive lactic acid bacteria possess a specific cell wall structure that allows for secretion of proteins directly into their environment. In spite of some limitations, this advantage and the fact that many of these bacteria have GRAS (generally recognized as safe) status made them into an attractive tool in biotechnology. In this chapter we give an overview of the potential of lactobacilli to be used for recombinant protein expression and their feasibility in various applications. In the past years lactobacilli have been used for the production of therapeutics, as vaccines, and as a platform for protein surface display. Many tools for genetic modification of lactobacilli have been developed and optimized. In addition, a great variety of gene regulatory elements have been isolated, characterized and established in the context of recombinant protein expression. We discuss the properties of different plasmids that may be used for the expression of intracellular or secreted target proteins. We describe the mechanisms of inducible and constitutive promoters, the influence of codon usage and ribosomal binding sites on protein expression levels, and strategies for stable integration of target genes into the genome of lactobacilli using modern technologies. Overall, there exists a high variety of different lactobacilli species and strains that are highly specialized and have adapted to specific ecological niches. Depending on the application, the desired product or the required process, genetic tools and regulatory elements often have to be tested, adapted and optimized individually read more ...
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