Heterologous expression of the lactacin F peptides by Carnobacterium piscicola LV17.
The lactacin F complex, composed of LafA and LafX peptides, is produced by Lactobacillus johnsonii VPI 11088 and is active against five other Lactobacillus species and Enterococcus faecalis. The genetic determinants encoding the lactacin F complex are organized in a 1-kb polycistronic operon which comprises three genes, lafA, lafX, and ORFZ (encoding the putative immunity protein). The lafA and lafX genes encode the bacteriocin precursors with N-terminal extensions characterized by a Gly-Gly-1*Xaa+1 cleavage site (*). The Gly-Gly motif is conserved in several other bacteriocins, including carnobacteriocins A, BM1, and B2. Carnobacterium piscicola LV17 produces carnobacteriocins which are active against Listeria monocytogenes and other lactic acid bacteria. In this study, the lactacin F operon was introduced into C. piscicola LV17. The transformants produced lactacin F concurrently with the carnobacteriocins. When the lafA and lafX genes were separated and cloned individually into LV17, production of either LafA or LafX by C. piscicola LV17 was detected by complementation with L. johnsonii clones producing LafX or LafA, respectively. Transformants of C. piscicola LV17 which produced lactacin F, LafA, or LafX, in combination with the carnobacteriocins, were assayed for an increased and expanded inhibitory spectrum. The recombinant organisms were only active against lactacin F- and carnobacteriocin-sensitive strains. A plasmidless derivative of LV17 which does not produce the carnobacteriocins failed to produce lactacin F, LafA, or LafX when transformed with the appropriate recombinant plasmids. The ability of C. piscicola LV17 to produce lactacin F demonstrates that the machinery for the carnobacteriocins is capable of processing and exporting bacteriocins from both systems.
Allison
G E
GE
Department of Microbiology, North Carolina State University, Raleigh 27695-7624, USA.
Worobo
R W
RW
Stiles
M E
ME
Klaenhammer
T R
TR
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D003001:Cloning, Molecular; D013293:Enterococcus faecalis / Q000187:drug effects; D005516:Food Microbiology; D005519:Food Preservation; D015870:Gene Expression; D005798:Genes, Bacterial; D006093:Gram-Positive Asporogenous Rods / Q000235:genetics* / Q000378:metabolism; D007778:Lactobacillus / Q000187:drug effects; D008969:Molecular Sequence Data; D009876:Operon; D021382:Protein Sorting Signals / Q000235:genetics; D014170:Transformation, Genetic
D001430:Bacteriocins; D021382:Protein Sorting Signals; C067573:lactacin F