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Production of Antilisterial Bacteriocins from Lactic Acid Bacteria in Dairy-Based Media: A Comparative Study

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Abstract

One hundred and eight strains of lactic acid bacteria (LAB) were screened for bacteriocin production by the modified deferred antagonism and agar well diffusion methods. When the modified deferred antagonism method was employed, 82 LAB strains showed inhibitory action against Listeria monocytogenes v7 ½a, whereas 26 LAB strains expressed no inhibition. Only 12 LAB strains exhibited inhibitory activity when the agar well diffusion method was used, 11 of which had been previously recognized as bacteriocin production positive (Bac+). Lactobacillus viridescens NRRL B-1951 was determined, for the first time, to produce an inhibitory compound with a proteinaceous nature. The inhibitory activity was observed in the presence of lipase, α-chymotrypsin, and trypsin, but no inhibition zone could be detected in the presence of proteinase K, indicating the proteinaceous nature of the inhibitory compound. The inhibitory compound was active against Lact. sake ATCC 15521 and Lact. plantarum NCDO 995. Bacteriocin production by the Bac+ LAB strains was assessed in Lactobacillus MRS Broth as well as in dairy-based media such as nonfat milk, demineralized whey powder, and cheddar cheese whey supplemented with complex nutrient sources that are rich in nitrogen. Lact. sake ATCC 15521 and L. monocytogenes CWD 1002, CWD 1092, CWD 1157, CWD 1198, and v7 ½a were used as indicators. The inhibitory activities of the bacteriocins varied depending on the indicator strains and the growth media used. The LAB indicator strains were found to be more sensitive to inhibition by bacteriocins when compared to the listerial indicator strains. Among the listerial indicators, L. monocytogenes CWD 1002 and CWD 1198 were the most sensitive strains to the bacteriocins investigated in this study. Media composition had a significant influence on bacteriocin production and activity. When compared to demineralized whey powder medium and cheddar cheese whey medium supplemented with whey protein concentrate, cheddar cheese whey medium supplemented with complex nutrient sources such as yeast extract, polypeptone, proteose peptone nr. 3, or soytone appeared to be more supportive of bacteriocin production.

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Acknowledgments

This project was supported by (1) The United States Department of Agriculture (USDSA) Cooperative State Research, Education, and Extension (CSREES) Hatch (IDA01369); (2) Idaho Beef Council (IBC); (3) United Dairymen of Idaho (UDI); and (4) MJ Murdock Charitable Trust. We thank all colleagues and institutions that provided us with bacterial strains. We also thank Dr. William Price of the College of Agricultural and Life Sciences (CALS) Statistical Services and Dr. Lee Deobald of the Department of Chemistry, both at the University of Idaho, for their assistance with statistical analyses and freeze-drying experiments, respectively.

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  1. Claudia Ionita

    Present address: Department of Food, Nutrition, and Packaging Sciences, Clemson University, 223 Poole Agricultural Center, Box 340316, Clemson, SC, 29634-0316, USA

Authors and Affiliations

  1. School of Food Science, University of Idaho, 875 Perimeter Drive, Moscow, ID, 83844-2312, USA

    Gülhan Ünlü, Barbara Nielsen & Claudia Ionita

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  1. Gülhan Ünlü
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Correspondence to Gülhan Ünlü.

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Gülhan Ünlü, Barbara Nielsen and Claudia Ionita declares that they have no conflict of interest.

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Ünlü, G., Nielsen, B. & Ionita, C. Production of Antilisterial Bacteriocins from Lactic Acid Bacteria in Dairy-Based Media: A Comparative Study. Probiotics & Antimicro. Prot. 7, 259–274 (2015). https://doi.org/10.1007/s12602-015-9200-z

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