Heterologous expression of the bacteriocin mesentericin Y105 using the dedicated transport system and the general secretion pathway.
Two different N-terminal extensions have been identified within class II bacteriocin precursors. The first one is a two-glycine-type leader peptide associated with a dedicated ATP-binding cassette transporter. The second is a signal peptide which directs the bacteriocin precursor to the general secretion machinery. Mesentericin Y105 is a class II anti-Listeria bacteriocin produced by Leuconostoc mesenteroides Y105 via a dedicated transport system (DTS). To investigate heterologous expression systems capable of producing mesentericin Y105 in various hosts, two different secretion vectors were constructed. One of them, containing the mesentericin Y105 structural gene fused to the segment encoding the divergicin A signal peptide, was introduced into Escherichia coli, Leuconostoc subsp. and Lactococcus subsp. In E. coli, mesentericin Y105 production was linked to a putative periplasmic toxicity. To take advantage of this secretion system, the mesentericin Y105 precursor was also produced in E. coli. It was demonstrated that this pre-bacteriocin exhibited some antagonistic activity against Listeria. To allow for a comparison between the two different transport systems, mesentericin Y105 production using the vector containing the mesentericin Y105 structural gene and its DTS transporter operon was examined. The production of mesentericin Y105 was monitored by a new fast purification method followed by MS analysis. It was shown that, in Leuconostoc, the production of mesentericin Y105 is enhanced via the DTS compared to the general secretion pathway.
Biet
Franck
F
lnstitut de Biologie MolCculaire et d'lnghierie GCnCtiqueCNRS-ESA 6031, UniversitC de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers CedexFrance.
Berjeaud
Jean Marc
JM
lnstitut de Biologie MolCculaire et d'lnghierie GCnCtiqueCNRS-ESA 6031, UniversitC de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers CedexFrance.
Worobo
Randy W
RW
lnstitut de Biologie MolCculaire et d'lnghierie GCnCtiqueCNRS-ESA 6031, UniversitC de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers CedexFrance.
Cenatiempo
Yves
Y
lnstitut de Biologie MolCculaire et d'lnghierie GCnCtiqueCNRS-ESA 6031, UniversitC de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers CedexFrance.
Fremaux
Christophe
C
Texel, groupe RhGne- PoulencZA de Buxieres BP 10, 86220 DangC Saint- RomainFrance.|lnstitut de Biologie MolCculaire et d'lnghierie GCnCtiqueCNRS-ESA 6031, UniversitC de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers CedexFrance.
D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D001430:Bacteriocins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001692:Biological Transport; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000235:genetics / Q000378:metabolism; D007934:Leuconostoc / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D011498:Protein Precursors / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D014169:Transformation, Bacterial
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C093812:dvnA protein, Carnobacterium divergens; C079275:mesentericin Y105 protein, Leuconostoc mesenteroides