Production of pediocin PA-1 by Lactococcus lactis using the lactococcin A secretory apparatus.
The class II bacteriocins pediocin PA-1, from Pediococcus acidilactici, and lactococcin A, from Lactococcus lactis subsp. lactis bv. diacetylactis WM4 have a number of features in common. They are produced as precursor peptides containing similar amino-terminal leader sequences with a conserved processing site (Gly-Gly at positions -1 and -2). Translocation of both bacteriocins occurs via a dedicated secretory system. Because of the strong antilisterial activity of pediocin PA-1, its production by lactic acid bacteria strains adapted to dairy environments would considerably extend its application in the dairy industry. In this study, the lactococcin A secretory system was adapted for the expression and secretion of pediocin PA-1. A vector containing an inframe fusion of sequences encoding the lcnA promoter, the lactococcin A leader, and the mature pediocin PA-1, was introduced into L. lactis IL1403. This strain is resistant to pediocin PA-1 and encodes a lactococcin translocation apparatus. The resulting L. lactis strains secreted a bacteriocin with an antimicrobial activity of approximately 25% of that displayed by the parental pediocin-producing P. acidilactici 347. A noncompetitive indirect enzyme-linked immunosorbent assay with pediocin PA-1-specific antibodies and amino-terminal amino acid sequencing confirmed that pediocin PA-1 was being produced by the heterologous host.
Horn
N
N
Department of Genetics and Microbiology, Institute of Food Research, Colney, Norwich, United Kingdom.
Martínez
M I
MI
Martínez
J M
JM
Hernández
P E
PE
Gasson
M J
MJ
Rodríguez
J M
JM
Dodd
H M
HM
D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001430:Bacteriocins / Q000096:biosynthesis / Q000302:isolation & purification; D013294:Lactococcus lactis / Q000378:metabolism*; D000070017:Pediocins; D011994:Recombinant Proteins / Q000096:biosynthesis*
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D000070017:Pediocins; D011994:Recombinant Proteins; C074288:pediocin PA-1