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Applied and Environmental Microbiology, October 2000, p. 4427-4432, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Adaptation of the Nisin-Controlled Expression System in Lactobacillus plantarum: a Tool To Study In Vivo Biological Effects

Sonia Pavan,1 Pascal Hols,2 Jean Delcour,2 Marie-Claude Geoffroy,1 Corinne Grangette,1 Michiel Kleerebezem,3 and Annick Mercenier1,*

Département de Microbiologie des Ecosystèmes, Institut Pasteur de Lille, F-59019 Lille Cedex, France1; Unité de Génétique, Université Catholique de Louvain, B-1348 Louvain-la-Neuve, Belgium2; and Wageningen Centre for Food Sciences, NIZO Food Research, 6710 BA Ede, The Netherlands3

Received 13 April 2000/Accepted 28 July 2000

The potential of lactic acid bacteria as live vehicles for the production and delivery of therapeutic molecules is being actively investigated today. For future applications it is essential to be able to establish dose-response curves for the targeted biological effect and thus to control the production of a heterologous biopeptide by a live lactobacillus. We therefore implemented in Lactobacillus plantarum NCIMB8826 the powerful nisin-controlled expression (NICE) system based on the autoregulatory properties of the bacteriocin nisin, which is produced by Lactococcus lactis. The original two-plasmid NICE system turned out to be poorly suited to L. plantarum. In order to obtain a stable and reproducible nisin dose-dependent synthesis of a reporter protein (beta -glucuronidase) or a model antigen (the C subunit of the tetanus toxin, TTFC), the lactococcal nisRK regulatory genes were integrated into the chromosome of L. plantarum NCIMB8826. Moreover, recombinant L. plantarum producing increasing amounts of TTFC was used to establish a dose-response curve after subcutaneous administration to mice. The induced serum immunoglobulin G response was correlated with the dose of antigen delivered by the live lactobacilli.

* Corresponding author. Mailing address: Département de Microbiologie des Ecosystèmes, Institut Pasteur de Lille, 1, rue du Pr. Calmette, B. P. 245, F-59019 Lille Cedex, France. Phone: (33) 320-87-71-22. Fax: (33) 320-87-79-08. E-mail: annick.mercenier{at}pasteur-lille.fr.

Applied and Environmental Microbiology, October 2000, p. 4427-4432, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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