title: Heterologous protein secretion in Lactobacilli with modified pSIP vectors.
abstract: We describe new variants of the modular pSIP-vectors for inducible gene expression and protein secretion in lactobacilli. The basic functionality of the pSIP system was tested in Lactobacillus strains representing 14 species using pSIP411, which harbors the broad-host-range Lactococcus lactis SH71rep replicon and a β-glucuronidase encoding reporter gene. In 10 species, the inducible gene expression system was functional. Based on these results, three pSIP vectors with different signal peptides were modified by replacing their narrow-host-range L. plantarum 256rep replicon with SH71rep and transformed into strains of five different species of Lactobacillus. All recombinant strains secreted the target protein NucA, albeit with varying production levels and secretion efficiencies. The Lp_3050 derived signal peptide generally resulted in the highest levels of secreted NucA. These modified pSIP vectors are useful tools for engineering a wide variety of Lactobacillus species.
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway.
authors.forename: Ingrid Lea
authors.initials: IL
authors.lastname: Karlskås
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway.
authors.forename: Kristina
authors.initials: K
authors.lastname: Maudal
authors.affiliation: Nofima - Norwegian Institute for Food, Fisheries and Aquaculture Research, Ås, Norway.
authors.forename: Lars
authors.initials: L
authors.lastname: Axelsson
authors.affiliation: Nofima - Norwegian Institute for Food, Fisheries and Aquaculture Research, Ås, Norway.
authors.forename: Ida
authors.initials: I
authors.lastname: Rud
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway.
authors.forename: Vincent G H
authors.initials: VG
authors.lastname: Eijsink
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway.
authors.forename: Geir
authors.initials: G
authors.lastname: Mathiesen
mesh_terms: D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D018628:Gene Dosage; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000378:metabolism*; D058507:Host Specificity; D007778:Lactobacillus / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D016622:Silver Staining; D013696:Temperature
chemical_list: D011994:Recombinant Proteins