Skip to main content

Advertisement

Springer Nature Link
Log in

Identification and in vitro assessment of potential probiotic characteristics and antibacterial effects of Lactobacillus plantarum subsp. plantarum SKI19, a bacteriocinogenic strain isolated from Thai fermented pork sausage

  • Original Article
  • Published:
Journal of Food Science and Technology Aims and scope Submit manuscript

Abstract

A total of 2257 lactic acid bacteria were preliminarily screened for antagonistic activity against Lactobacillus sakei subsp. sakei JCM 1157. Strain SKI19 was selected and identified at the subspecies level as Lactobacillus plantarum subsp. plantarum SKI19, using 16S rRNA gene sequence analysis combined with recA and dnaK genes’ amplification. Antibacterial activity of SKI19 was completely lost after treatment of neutralized cell free culture supernatant with proteolytic enzymes, suggesting that SKI19 produced a bacteriocin-like substance that inhibited not only closely related species, but was also effective against Listeria monocytogenes DMST 17303. Viewed under scanning electron microscope, cell membranes of the indicator strain appeared to collapse after exposure to the bacteriocin-like substance. In vitro tests concerning probiotic properties, SKI19 survived under simulated gastrointestinal tract conditions, and adhesion of its cell surface to xylene and chloroform was 90.14 and 89.85%, respectively. Complete inhibition by SKI19 against pathogenic bacteria (Escherichia coli DMST 4212, L. monocytogenes DMST 17303, and Staphylococcus aureus DMST 8840) was observed in co-cultivation under anaerobic conditions. A safety assessment showed that SKI19 was susceptible to several antibiotics and had no haemolytic activity. PCR amplification of virulence factors with the specific primers for ace, asa1, cylLS, efaAfs, hyl, and gelE genes were negative for SKI19. Also, SKI19 did not harbor any hdc, tdc, odc or ldc genes involved in biogenic amine production. The results reveal that SKI19 has probiotic potential and antibacterial activity, and is safe for further application in certain food products.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Aslim B, Yuksekda ZN, Sarikaya E, Beyatli Y (2005) Determination of the bacteriocin-like substances produced by some lactic acid bacteria isolated from Turkish dairy products. LWT Food Sci Technol 38:691–694

    Article  CAS  Google Scholar 

  • Balcázar JL, Decamp O, Vendrell D, Blas ID, Ruiz-Zarzuela I (2006) Health and nutritional properties of probiotics in fish and shellfish. Microb Ecol Health Dis 18:65–70

    Article  CAS  Google Scholar 

  • Barbosa MS, Todorov SD, Belguesmia Y, Choiset Y, Rabesona H, Ivanova IV, Chobert JM, Haertlé T, Franco BDGM (2014) Purification and characterization of the bacteriocin produced by Lactobacillus sakei MBSa1 isolated from Brazilian salami. J Appl Microbiol 116:1195–1208

    Article  CAS  PubMed  Google Scholar 

  • Belicova A, Mikulasova M, Dusinsky M (2013) Probiotic potential and safety properties of Lactobacillus plantarum from Slovak bryndza cheese. Biomed Res Int. https://doi.org/10.1155/2013/760298

    Article  PubMed  PubMed Central  Google Scholar 

  • Cleveland J, Montville TJ, Nes IF, Chikindas ML (2001) Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 71:1–20

    Article  CAS  PubMed  Google Scholar 

  • de las Rivas B, Marcobal A, Carrascosa AV, Muñoz R (2006) PCR detection of foodborne bacteria producing the biogenic amines histamine, tyramine, putrescine, and cadaverine. J Food Prot 69:2509–2514

    Article  PubMed  Google Scholar 

  • Eaton TJ, Gasson MJ (2001) Molecular screening of Enterococcus virulence determinants and potential for genetic exchange between food and medical isolates. Appl Environ Microbiol 67:1628–1635

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • EFSA (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J 10:2740

    Google Scholar 

  • FAO/WHO (2002) Guidelines for the evaluation of probiotics in food. London, Ontario, Canada, April 30 and May 1. http://www.who.int/foodsafety/fs_management/en/probiotic_guidelines.pdf. Accessed 16 July 2017

  • Fraqueza MJ (2015) Antibiotic resistance of lactic acid bacteria isolated from dry-fermented sausages. Int J Food Microbiol 212:76–88

    Article  CAS  PubMed  Google Scholar 

  • Gobbetti M, Di Cagno M, De Angelis M (2010) Functional microorganisms for functional food quality. Crit Rev Food Sci Nutr 50:716–727

    Article  CAS  PubMed  Google Scholar 

  • Gong HS, Meng XC, Wang H (2010) Plantaricin MG active against Gram-negative bacteria produced by Lactobacillus plantarum KLDS1.0391 isolated from “Jiaoke”, a traditional fermented cream from China. Food Control 21:89–96

    Article  CAS  Google Scholar 

  • H-Kittikun A, Biscola V, El-Ghaish S, Jaffrès E, Dousset X, Pillot G, Haertlé T, Chobert JM, Hwanhlem N (2015) Bacteriocin-producing Enterococcus faecalis KT2W2G isolated from mangrove forests in southern Thailand: purification, characterization and safety evaluation. Food Control 54:126–134

    Article  CAS  Google Scholar 

  • Hongpattarakere T, Cherntong N, Wichienchot S, Kolida S, Rastall RA (2012) In vitro prebiotic evaluation of exopolysaccharides produced by marine isolated lactic acid bacteria. Carbohydr Polym 87:846–852

    Article  CAS  Google Scholar 

  • Huang Y, Adams MC (2004) In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria. Int J Food Microbiol 91:253–260

    Article  PubMed  Google Scholar 

  • Huang CH, Lee FL, Liou JS (2010) Rapid discrimination and classification of the Lactobacillus plantarum group based on a partial dnaK sequence and DNA fingerprinting techniques. Antonie Van Leeuwenhoek 97:289–296

    Article  CAS  PubMed  Google Scholar 

  • Hwanhlem N, Ivanova T, Biscola V, Choiset Y, Haertlé T (2017) Bacteriocin producing Enterococcus faecalis isolated from chicken gastrointestinal tract originating from Phitsanulok, Thailand: isolation, screening, safety evaluation and probiotic properties. Food Control 78:187–195

    Article  CAS  Google Scholar 

  • Kaushik JK, Kumar A, Duary RK, Mohanty AK, Grover S, Batish VK (2009) Functional and probiotic attributes of an indigenous isolate of Lactobacillus plantarum. PLoS ONE. https://doi.org/10.1371/journal.pone.0008099

    Article  PubMed  PubMed Central  Google Scholar 

  • Khan I, Kang SC (2016) Probiotic potential of nutritionally improved Lactobacillus plantarum DGK-17 isolated from Kimchi—a traditional Korean fermented food. Food Control 60:88–94

    Article  CAS  Google Scholar 

  • Kongnum K, Hongpattarakere T (2012) Effect of Lactobacillus plantarum isolated from digestive tract of wild shrimp on growth and survival of white shrimp (Litopenaeus vannamei) challenged with Vibrio harveyi. Fish Shellfish Immunol 32:170–177

    Article  PubMed  Google Scholar 

  • Landeta G, Curiel JA, Carrascosa AV, Munõz R, de las Rivas B (2013) Technological and safety properties of lactic acid bacteria isolated from Spanish dry-cured sausages. Meat Sci 95:272–280

    Article  CAS  PubMed  Google Scholar 

  • Maldonado A, Ruiz-Barba JL, Jiménez-Díaz R (2003) Purification and genetic characterization of plantaricin NC8, a novel co-culture-inducible two-peptide bacteriocin from Lactobacillus plantarum NC8. Appl Environ Microbiol 69:383–389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martín-Platero AM, Valdivia E, Maqueda M, Martínez-Bueno M (2009) Characterization and safety evaluation of enterococci isolated from Spanish goats’ milk cheeses. Int J Food Microbiol 132:24–32

    Article  CAS  PubMed  Google Scholar 

  • Oh YJ, Jung DS (2015) Evaluation of probiotic properties of Lactobacillus and Pediococcus strains isolated from Omegisool, a traditionally fermented millet alcoholic beverage in Korea. LWT Food Sci Technol 63:437–444

    Article  CAS  Google Scholar 

  • Paukatong KV, Kunawasen S (2001) The Hazard Analysis and Critical Control Points (HACCP) generic model for the production of Thai fermented pork sausage (Nham). Berl Munch Tierarztl Wochenschr 114:327–330

    CAS  PubMed  Google Scholar 

  • Perin LM, Miranda RO, Todorov SD, de Melo Franco BDG, Nero LA (2014) Virulence, antibiotic resistance and biogenic amines of bacteriocinogenic lactococci and enterococci isolated from goat milk. Int J Food Microbiol 185:121–126

    Article  CAS  PubMed  Google Scholar 

  • Pieniz S, de Moura TM, Cassenego APV, Andreazza R, Frazzon APG, de Oliveira Camargo FA, Brandelli A (2015) Evaluation of resistance genes and virulence factors in a food isolated Enterococcus durans with potential probiotic effect. Food Control 51:49–54

    Article  CAS  Google Scholar 

  • Powell JE, Witthuhn RC, Todorov SD, Dicks LMT (2007) Characterization of bacteriocin ST8KF produced by a kefir isolate Lactobacillus plantarum ST8KF. Int Dairy J 17:190–198

    Article  CAS  Google Scholar 

  • Semedo T, Santos MA, Martins P, Lopes MFS, Marques JJF, Tenreiro R, Crespo MTB (2003) Comparative study using type strains and clinical and food isolates to examine hemolytic activity and occurrence of the cyl operon in enterococci. J Clin Microbiol 41:2569–2576

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Swetwiwathana A, Visessanguan W (2015) Potential of bacteriocin-producing lactic acid bacteria for safety improvements of traditional Thai fermented meat and human health. Meat Sci 109:101–105

    Article  CAS  PubMed  Google Scholar 

  • Szala B, Paluszak Z, Motyl I (2012) Antagonistic effect of Lactic acid bacteria on Salmonella Senftenberg in mixed cultures. Pol J Environ Stud 21:1399–1403

    Google Scholar 

  • Torriani S, Felis GE, Dellaglio F (2001) Differentiation of Lactobacillus plantarum, L. pentosus, and L. paraplantarum by recA gene sequence analysis and multiplex PCR assay with recA gene-derived primers. Appl Environ Microbiol 67:3450–3454

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vankerckhoven V, Huys G, Vancanneyt M, Vael C, Klare I, Romond MB, Entenza JM, Moreillon P, Wind RD, Knol J, Wiertz E, Pot B, Vaughan EE, Kahlmeter G, Goossens H (2008) Biosafety assessment of probiotics used for human consumption: recommendations from the EU-PROSAFE project. Trends Food Sci Technol 19:102–114

    Article  CAS  Google Scholar 

  • Visessanguan W, Plengvidhya V, Chokesajjawatee N, Bakar JA (2015) Lactic meat fermentation. In: Owens JD (ed) Indigenous fermented foods of Southeast Asia. CRC Press, New York, pp 313–358

    Google Scholar 

  • Vuyst LD, Falony G, Leroy F (2008) Probiotics in fermented sausages. Meat Sci 80:75–78

    Article  CAS  PubMed  Google Scholar 

  • Yu Z, Zhang X, Li S, Li C, Li D, Yang Z (2013) Evaluation of probiotic properties of Lactobacillus plantarum strains isolated from Chinese sauerkraut. World J Microbiol Biotechnol 29:489–498

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Higher Education Research Promotion and the Thailand’s Education Hub for Southern Region of ASEAN Countries Project Office of the Higher Education Commission. Also, the research was funded by the Graduate School, Prince of Songkla University, the Thailand Research Fund and the Commission on Higher Education for Project No. MRG5080138, and the Research and Development Office, Prince of Songkla University. The authors would like to thank Associate Professor Seppo J. Karrila, Ph.D. (Chem Eng) for the professional English proofreading service.

Author information

Authors and Affiliations

  1. Department of Industrial Biotechnology, Faculty of Agro-Industry, Prince of Songkla University, Songkhla, 90112, Thailand

    Viengvilaiphone Botthoulath, Apichat Upaichit & Uschara Thumarat

Authors
  1. Viengvilaiphone Botthoulath
    View author publications

    Search author on:PubMed Google Scholar

  2. Apichat Upaichit
    View author publications

    Search author on:PubMed Google Scholar

  3. Uschara Thumarat
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Apichat Upaichit.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 16 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Botthoulath, V., Upaichit, A. & Thumarat, U. Identification and in vitro assessment of potential probiotic characteristics and antibacterial effects of Lactobacillus plantarum subsp. plantarum SKI19, a bacteriocinogenic strain isolated from Thai fermented pork sausage. J Food Sci Technol 55, 2774–2785 (2018). https://doi.org/10.1007/s13197-018-3201-3

Download citation

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13197-018-3201-3

Keywords