Premium
High‐level gene expression in Lactobacillus plantarum using a pheromone‐regulated bacteriocin promoter
Author(s) -
Mathiesen G.,
Sørvig E.,
Blatny J.,
Naterstad K.,
Axelsson L.,
Eijsink V.G.H.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01551.x
Subject(s) - biology , lactococcus lactis , gene , operon , plasmid , bacteriocin , lactobacillus plantarum , gene expression , reporter gene , promoter , regulator gene , regulation of gene expression , escherichia coli , genetics , bacteria , lactic acid
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.