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Appl Environ Microbiol, March 1998, p. 818-823, Vol. 64, No. 3
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Production of Pediocin PA-1 by Lactococcus lactis Using the Lactococcin A Secretory Apparatus

Nikki Horn,1 María I. Martínez,2 José M. Martínez,2 Pablo E. Hernández,2 Michael J. Gasson,1 Juan M. Rodríguez,2 and Helen M. Dodd1,*

Department of Genetics and Microbiology, Institute of Food Research, Colney, Norwich NR4 7UA, United Kingdom,1 and Departamento de Nutrición y Bromatología III, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain2

Received 15 October 1997/Accepted 19 December 1997

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 in-frame 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.

* Corresponding author. Mailing address: Department of Genetics and Microbiology, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. Phone: 44 1603 255243. Fax: 44 1603 507723. E-mail: HELEN.DODD{at}BBSRC.AC.UK.




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