High-level gene expression in Lactobacillus plantarum using a pheromone-regulated bacteriocin promoter.
OBJECTIVE
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.


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 beta-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.


CONCLUSIONS
This study describes a system for controllable gene expression in lactobacilli, giving some of the highest expression levels reported so far in this genus.
Mathiesen
G
G
Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, As, Norway.
Sørvig
E
E
Blatny
J
J
Naterstad
K
K
Axelsson
L
L
Eijsink
V G H
VG
D003086:Bacteriocin Plasmids / Q000235:genetics / Q000378:metabolism; D001430:Bacteriocins / Q000235:genetics*; D018826:CD13 Antigens / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005966:Glucuronidase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D007778:Lactobacillus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics; D010675:Pheromones / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics
D001430:Bacteriocins; D010675:Pheromones; D011993:Recombinant Fusion Proteins; C087217:sakacin P protein, Lactobacillus; D005966:Glucuronidase; D018826:CD13 Antigens