Chimeras of mature pediocin PA-1 fused to the signal peptide of enterocin P permits the cloning, production, and expression of pediocin PA-1 in Lactococcus lactis.
Chimeras of pediocin PA-1 (PedA-1), a bacteriocin produced by Pediococcus acidilactici PLBH9, fused to the signal peptide of enterocin P (EntP), a sec-dependent bacteriocin produced by Enterococcus faecium P13, permitted the production of PedA-1 in Lactococcus lactis. Chimeric genes encoding the EntP signal peptide (SP(entP)) fused to mature PedA-1 (pedA), with or without its immunity gene (pedB), were cloned into the expression vector pMG36c to generate the recombinant plasmids pMPP9 (SP(entP):pedA) and pMPP14i (SP(entP):pedA + pedB). Transformation of competent L. lactis subsp. lactis IL1403, L. lactis subsp. cremoris NZ9000, and L. lactis subsp. lactis DPC5598 with the recombinant plasmids has permitted the detection and quantitation of PedA-1 and the coproduction of nisin A and PedA-1 in supernatants of producer cells with specific anti-PedA-1 antibodies and a noncompetitive indirect enzyme-linked immunosorbent assay. Recombinant L. lactis hosts carrying pMPP9 or pMPP14i displayed antimicrobial activity, suggesting that mature PedA-1 fused to SP(EntP) is the minimum requirement for the synthesis, processing, and secretion of biologically active PedA-1 in L. lactis. However, the production and antimicrobial activity of the PedA-1 produced by L. lactis was lower than that produced by the P. acidilactici control strains.
Martín
María
M
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain.
Gutiérrez
Jorge
J
Criado
Raquel
R
Herranz
Carmen
C
Cintas
Luis M
LM
Hernández
Pablo E
PE
D001430:Bacteriocins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics*; D002678:Chimera; D003001:Cloning, Molecular; D016984:Enterococcus faecium / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D013294:Lactococcus lactis / Q000737:chemistry* / Q000235:genetics*; D009561:Nisin / Q000096:biosynthesis; D000070017:Pediocins; D010376:Pediococcus / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D021382:Protein Sorting Signals / Q000502:physiology
D001430:Bacteriocins; D000070017:Pediocins; D021382:Protein Sorting Signals; C417717:enterocin P; C074288:pediocin PA-1; D009561:Nisin; C102526:nisin A