Inhibitory Effect of Substances Produced by Native Lactococcus lactis Strains of Tropical Fruits towards Food Pathogens

다국어 초록 (Multilingual Abstract)

The use of peptides produced by lactic acid bacteria (LAB) as antimicrobial agents in food emerged from the increasing need of replacing chemicals with natural substances to ensure their safety and quality. A total of 30 LAB belonging to the genus Lactococcus sp. (10) and Enterococcus sp. (20) were isolated from native fruits of Ecuador subtropical rainforest. Among Lactococcus species, the isolates assigned Gt28, Gt29, and Ella8, identified as Lactococcus lactis subsp. lactis with 99% identity, showing highly inhibitory potential against four food pathogens were further characterized. The treatment of cell-free supernatant with proteolytic enzymes indicated the protein nature of released components, which displayed a broad antimicrobial activity against Gram-positive and -negative bacteria. Polymerase chain reaction analysis indicated the presence of lacticin 3147 gene in all isolates, lactococcin M gene in Gt28 and Gt29 but not in Ella8 and lactococcin A gene in Gt28 only. The antimicrobial activity was not linked to the presence of structural nisin gene as no amplification product was obtained. Treatment of Salmonella enterica ATCC 51741 and Escherichia coli ATCC 25922 at both vegetative and exponential phase of growth with the cell-free supernatant of Gt28 resulted in complete inactivation upon 3 h suggesting its bactericidal mode of action. An increment on inhibitory activity occurred when partial purified bacteriocin Gt28 was combined with ethylenediaminetetraacetic acid rather than bacteriocin only, indicating that the cells were sensitized in vitro by the chelator agent acting synergistically to induce the killing of pathogenic cells.

목차 (Table of Contents)

• ABSTRACT
• INTRODUCTION
• MATERIALS AND METHODS
• RESULTS AND DISCUSSION
• REFERENCES

참고문헌 (Reference)

1 Moschetti G, "sacA and nisA genes are not always linked in Lactococcus lactis subsp. lactis strains" 170 : 373-379, 1999

2 Arena MP, "Use of Lactobacillus plantarum strains as a bio-control strategy against food-borne pathogenic microorganisms" 7 : 464-, 2016

3 Draper LA, "The two peptide lantibiotic lacticin 3147 acts synergistically with polymyxin to inhibit Gram negative bacteria" 13 : 212-, 2013

4 European Food Safety Authority, "The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2015" 14 : 4634-, 2016

5 Chopra L, "Sonorensin: a new bacteriocin with potential of an anti-biofilm agent and a food biopreservative" 5 : 13422-, 2015

6 Ge J, "Purification and partial characterization of a novel bacteriocin synthesized by Lactobacillus paracasei HD1-7 isolated from Chinese sauerkraut juice" 6 : 19366-, 2016

7 Rumjuankiat K, "Purification and characterization of a novel plantaricin, KL-1Y, from Lactobacillus plantarum KL-1" 31 : 983-994, 2015

8 Todorov SD, "Partial characterization of bacteriocins produced by three strains of Lactobacillus sakei, isolated from salpicao, a fermented meat product from North-West of Portugal" 30 : 111-121, 2013

9 Fontana C, "Occurrence of antilisterial structural bacteriocins genes in meat borne lactic acid bacteria" 47 : 53-59, 2015

10 Balciunas EM, "Novel biotechnological applications of bacteriocins: a review" 32 : 134-142, 2013

11 Gharsallaoui A, "Nisin as a food preservative. Part 1: physicochemical properties, antimicrobial activity, and main uses" 56 : 1262-1274, 2016

12 Delves-Broughton J, "Nisin as a food preservative" 57 : 525-527, 2005

13 Deraz SF, "Mode of action of acidocin D20079, a bacteriocin produced by the potential probiotic strain, Lactobacillus acidophilus DSM 20079" 34 : 34373-34379, 2007

14 NTE INEN, "Meat and meat products-sampling. Norma Tecnica Ecuatoriana, Instituto Ecuatoriano de Normalizacion, Quito, Ecuador" 2012

15 Ministerio de Salud Publica, "MSP lidera la primera campana de capacitacion en manipulacion de alimentos"

16 Suda S, "Lacticin 3147-biosynthesis, molecular analysis, immunity, bioengineering and applications" 13 : 193-204, 2012

17 Waterhouse AM, "Jalview version 2-a multiple sequence alignment editor and analysis workbench" 25 : 1189-1191, 2009

18 Noonpakdee W, "Isolation of nisin-producing Lactococcus lactis WNC 20 strain from nham, a traditional Thai fermented sausage" 81 : 137-145, 2003

19 Biscola V, "Isolation and characterization of a nisinlike bacteriocin produced by a Lactococcus lactis strain isolated from charqui, a Brazilian fermented, salted and dried meat product" 93 : 607-613, 2013

20 Li H, "Heterologous expression of the Lactococcus lactis bacteriocin, nisin, in a dairy Enterococcus strain" 68 : 3392-3400, 2002

21 Luche S, "Evaluation of nonionic and zwitterionic detergents as membrane protein solubilizers in two-dimensional electrophoresis" 3 : 249-253, 2002

22 Bhatia S, "Evaluating the antimicrobial activity of nisin, lysozyme and ethylenediaminetetraacetate incorporated in starch based active food packaging film" 52 : 3504-3512, 2015

23 Hartmann HA, "Efficacy of bacteriocin-containing cell-free culture supernatants from lactic acid bacteria to control Listeria monocytogenes in food" 146 : 192-199, 2011

24 Suda S, "Effect of bioengineering lacticin 3147 lanthionine bridges on specific activity and resistance to heat and proteases" 17 : 1151-1160, 2010

25 Ivanova I, "Detection, purification and partial characterization of a novel bacteriocin substance produced by Lactoccous lactis subsp. lactis B14 isolated from boza-Bulgarian traditional cereal beverage" 41 : 47-53, 2000

26 Banerjee SP, "Detection, partial purification and characterization of bacteriocin produced by Lactobacillus brevis FPTLB3 isolated from freshwater fish: bacteriocin from Lb. brevis FPTLB3" 50 : 17-25, 2013

27 Martinez-Cuesta MC, "Control of late blowing in cheese by adding lacticin 3147-producing Lactococcus lactis IFPL 3593 to the starter" 20 : 18-24, 2010

28 Garzon K, "Characterization of bacteriocin-producing lactic acid bacteria isolated from native fruits of Ecuadorian Amazon" 66 : 473-481, 2017

29 Fatima D, "Characterization and determination of the factors affecting anti-listerial bacteriocins from Lactobacillus plantarum and Pediococcus pentosaceus isolated from dairy milk products" 7 : 35-44, 2013

30 Maina JW, "Bacteriocins: limiting factors to optimum activity" 5 : 19-25, 2017

31 Deegan LH, "Bacteriocins: biological tools for bio-preservation and shelf-life extension" 16 : 1058-1071, 2006

32 Alegria A, "Bacteriocins produced by wild Lactococcus lactis strains isolated from traditional, starter-free cheeses made of raw milk" 143 : 61-66, 2010

33 Alvarez-Sieiro P, "Bacteriocins of lactic acid bacteria: extending the family" 100 : 2939-2951, 2016

34 Parada JL, "Bacteriocins from lactic acid bacteria: purification, properties and use as biopreservatives" 50 : 512-542, 2007

35 Hwanhlem N, "Bacteriocin-producing lactic acid bacteria isolated from mangrove forests in southern Thailand as potential bio-control agents: purification and characterization of bacteriocin produced by Lactococcus lactis subsp. lactis KT2W2L" 5 : 264-278, 2013

36 Benavides AB, "Assessment of the in vitro bioactive properties of lactic acid bacteria isolated from native ecological niches of Ecuador" 48 : 236-244, 2016

37 Pratush A, "Application of purified bacteriocin produced by Lactococcus lactis AP2 as food biopreservative in acidic foods" 13 : 82-87, 2012

38 El-Ghaish S, "Antimicrobial impact for Lactococcus lactis subsp. lactis A15 and Enterococcus faecium A15 isolated from some traditional Egyptian dairy products on some pathogenic bacteria" 41 : e12279-, 2017

39 Yang SC, "Antibacterial activities of bacteriocins: application in foods and pharmaceuticals" 5 : 241-, 2014

40 Coconnier MH, "Antagonistic activity of Lactobacillus acidophilus LB against intracellular Salmonella enterica serovar Typhimurium infecting human enterocyte-like Caco-2/TC-7 cells" 66 : 1152-1157, 2000