Present address: Janet Blatny, Norwegian Defence Research Establishment, PO Box 25, N-2027 Kjeller, Norway.
High-level gene expression in Lactobacillus plantarum using a pheromone-regulated bacteriocin promoter
Article first published online: 7 JUN 2004
Letters in Applied Microbiology
Volume 39, Issue 2, pages 137–143, August 2004
How to Cite
Mathiesen, G., Sørvig, E., Blatny, J., Naterstad, K., Axelsson, L. and Eijsink, V.G.H. (2004), High-level gene expression in Lactobacillus plantarum using a pheromone-regulated bacteriocin promoter. Letters in Applied Microbiology, 39: 137–143. doi: 10.1111/j.1472-765X.2004.01551.x
- Issue published online: 7 JUN 2004
- Article first published online: 7 JUN 2004
- 2004/0108: received 2 February 2004, revised 1 April 2004 and accepted 5 April 2004
- aminopeptidase N;
- Lactobacillus plantarum;
- sakacin P
Aims: To use promoters and regulatory genes involved in the production of the bacteriocin sakacin P to obtain high-level regulated gene expression in Lactobacillus plantarum.
Methods and Results: In a plasmid containing all three operons naturally involved in sakacin P production, the genes encoding sakacin P and its immunity protein were replaced by the aminopeptidase N gene from Lactococcus lactis (pepN) or the β-glucuronidase gene from Escherichia coli (gusA). The new genes were precisely fused to the start codon of the sakacin P gene and the stop codon of the immunity gene. This set-up permitted regulated (external pheromone controlled) overexpression of both reporter genes in L. plantarum NC8. For PepN, production levels amounted to as much as 40% of total cellular protein.
Conclusions: Promoters and regulatory genes involved in production of sakacin P are suitable for establishing inducible high-level gene expression in L. plantarum.
Significance and Impact of the Study: This study describes a system for controllable gene expression in lactobacilli, giving some of the highest expression levels reported so far in this genus.