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Applied and Environmental Microbiology, February 1999, p. 618-625, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Antilisterial Activity of Peptide AS-48 and Study of Changes Induced in the Cell Envelope Properties of an AS-48-Adapted Strain of Listeria monocytogenes

Fátima Mendoza,1 Mercedes Maqueda,2 Antonio Gálvez,3 Manuel Martínez-Bueno,1 and Eva Valdivia1,*

Departamento de Microbiología, Facultad de Ciencias,1 and Instituto de Biotecnología,2 Universidad de Granada, Granada, and Departamento de Microbiología, Facultad de Ciencias Experimentales, Universidad de Jaén, Jaén,3 Spain

Received 15 May 1998/Accepted 5 November 1998

The peptide AS-48 is highly active on all Listeria species. It has a bactericidal and bacteriolytic mode of action on Listeria monocytogenes CECT 4032, causing depletion of the membrane electrical potential and pH gradient. The producer strain Enterococcus faecalis A-48-32, releases sufficient amounts of AS-48 into the growth medium to suppress L. monocytogenes in cocultures at enterococcus-to-listeria ratios above 1 at 37°C or above 10 at 15°C. As the temperature decreases, the bactericidal effects of AS-48 are less pronounced, but at 2.5 µg/ml it still can inhibit the growth of listeria at 6°C. AS-48 is highly active on liquid cultures, although concentrations above 0.2 µg/ml are required to avoid adaptation of listeria. AS-48-adapted cells can be selected at low (but still inhibitory) concentrations, and they can be inhibited completely by AS-48 at 0.5 µg/ml. The adaptation is lost gradually upon repeated subcultivation. AS48ad cells are cross-resistant to nisin and show an increased resistance to muramidases. Their fatty acid composition is modified: they show a much higher proportion of branched fatty acids as well as a higher C15:0 An-to-C17:0 An ratio. Resistance to AS-48 is also maintained by protoplasts from AS48ad cells. Electron microscopy observations show that the cell wall of AS48ad cells is thicker and less dense. The structure of wild-type cells is severely modified after AS-48 treatment: the cell wall and the cytoplasmic membrane are disorganized, and the cytoplasmic content is lost. Intracytoplasmic membrane vesicles are also observed when the wild-type strain is treated with high AS-48 concentrations.

* Corresponding author. Mailing address: Departamento de Microbiología, Facultad de Ciencias, C/Fuentenueva s/n, 18071 Granada, Spain. Phone: 34-58-243244. Fax: 34-58-243246. E-mail: evavm{at}goliat.ugr.es.

Applied and Environmental Microbiology, February 1999, p. 618-625, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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