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Biomolecules 2015, 5(2), 1178-1194; doi:10.3390/biom5021178

A Microplate Growth Inhibition Assay for Screening Bacteriocins against Listeria monocytogenes to Differentiate Their Mode-of-Action

1
Department of Animal Science, Oklahoma State University, Monroe Street, Stillwater, OK 74078, USA
2
Robert M. Kerr Food & Agricultural Products Centre, Oklahoma State University, 109 FAPC Building, Monroe Street, Stillwater, OK 74078-6055, USA
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Jürg Bähler
Received: 6 March 2015 / Revised: 26 May 2015 / Accepted: 4 June 2015 / Published: 11 June 2015
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Abstract

Lactic acid bacteria (LAB) have historically been used in food fermentations to preserve foods and are generally-recognized-as-safe (GRAS) by the FDA for use as food ingredients. In addition to lactic acid; some strains also produce bacteriocins that have been proposed for use as food preservatives. In this study we examined the inhibition of Listeria monocytogenes 39-2 by neutralized and non-neutralized bacteriocin preparations (Bac+ preps) produced by Lactobacillus curvatus FS47; Lb. curvatus Beef3; Pediococcus acidilactici Bac3; Lactococcus lactis FLS1; Enterococcus faecium FS56-1; and Enterococcus thailandicus FS92. Activity differences between non-neutralized and neutralized Bac+ preps in agar spot assays could not readily be attributed to acid because a bacteriocin-negative control strain was not inhibitory to Listeria in these assays. When neutralized and non-neutralized Bac+ preps were used in microplate growth inhibition assays against L. monocytogenes 39-2 we observed some differences attributed to acid inhibition. A microplate growth inhibition assay was used to compare inhibitory reactions of wild-type and bacteriocin-resistant variants of L. monocytogenes to differentiate bacteriocins with different modes-of-action (MOA) whereby curvaticins FS47 and Beef3, and pediocin Bac3 were categorized to be in MOA1; enterocins FS92 and FS56-1 in MOA2; and lacticin FLS1 in MOA3. The microplate bacteriocin MOA assay establishes a platform to evaluate the best combination of bacteriocin preparations for use in food applications as biopreservatives against L. monocytogenes. View Full-Text
Keywords: bacteriocins; Listeria monocytogenes; microplate assay; inhibition; biopreservatives bacteriocins; Listeria monocytogenes; microplate assay; inhibition; biopreservatives
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Vijayakumar, P.P.; Muriana, P.M. A Microplate Growth Inhibition Assay for Screening Bacteriocins against Listeria monocytogenes to Differentiate Their Mode-of-Action. Biomolecules 2015, 5, 1178-1194.

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