Mikrobiol. Z. 2021; 83(1):12-20.
doi: https://doi.org/10.15407/microbiolj83.01.012

Effect of Cocultivation on Lactobacillus plantarum Strains Growth and Antagonistic Activity

I.L. Garmasheva, O.M. Vasyliuk, L.T. Oleschenko

Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

The use of bacterial starters for the production of fermented foods has several advantages over traditional spontaneous fermentation, as it provides a rapid and controlled decrease of pH, improves the microbiological quality of the product, and prolongs the shelf-life. Fermented foods are typically produced using mixed cultures of lactic acid bacteria (LAB) due to the synergism between their constituent bacterial cultures. So, the compatibility of the LAB strains decides the efficacy of a multi-strain starter. The purpose of this study was to investigate the effect of the cocultivation of Lactobacillus plantarum strains on the growth, acidification, and antagonistic activity to determine suitable strain combinations for fermented vegetable production. Methods. The effect of cocultivation on growth characteristics of four L. plantarum strains was determined in MRS medium and cabbage-based medium with 2.5% NaCl. After 8 h of cultivation at 30°C and 37°C, the number of viable cells (CFU/ml) and the pH of the medium were determined. The antagonistic activity of monocultures of L. plantarum and their six compositions against opportunistic pathogenic microorganisms was determined by the method of delayed antagonism. Results. During growth in MRS broth at 30°C cocultivation of L. plantarum 47SM with L. plantarum 691T or L. plantarum 1047K strains led to enhanced rates of growth compared to the monocultures, suggesting some degree of symbiosis between these strains. Viable cell counts of L. plantarum 47SM, 1047K and 691T strains and ΔpH values of L. plantarum 952K, 1047K, and 691T strains were higher after 8 h growth in the cabbage-based medium at 30°C compared to MRS broth. Despite the intensive growth of L. plantarum monocultures in cabbage-based medium, a significant decrease of viable cell counts and ΔpH values during cocultivation at 30°C were found. Cocultivation did not affect the average size of the growth inhibition zones of most of the indicator strains used. However, growth inhibition zones of Shigella flexneri, Escherichia coli, and Proteus vulgaris decreased in some L. plantarum mixed cultures compared to monocultures. Thus, the growth inhibition zones of E. coli and S. flexneri by mixed culture L. plantarum 47 SM+1047K were significantly smaller compared to the growth inhibition zones of L. plantarum monocultures. Conclusions. Thus, based on the data obtained in present work, we can assume that some of these L. plantarum strains used in the work may be bactericinogenic. Although the four L. plantarum strains studied are compatible when cocultivated in a standard rich MRS medium, the results of cocultivation in a cabbage-based medium with 2.5% NaCl does not allow to recommend the use of these L. plantarum strains simultaneously in the starter for vegetable fermentation. Further investigation of bacteriocinogenic properties and mechanisms of growth inhibition under cocultivation in vegetable-like conditions are needed, which will allow combining of some of these L. plantarum strains with LAB strains of other species or genera to create multi-starters for vegetable fermentation.

Keywords: cocultivation, vegetable fermentation, lactic acid bacteria, antagonism.

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  1. Şanlier N, Gökcen BB, Sezgin AC. Health benefits of fermented foods. Crit Rev Food Sci Nutr. 2019; 59(3):506–527. https://doi.org/10.1080/10408398.2017.1383355
  2. Bonatsou S, Tassou CC, Panagou EZ, Nychas GE. Table Olive Fermentation Using Starter Cultures with Multifunctional Potential. Microorganisms. 2017; 5(2):E30. https://doi.org/10.3390/microorganisms5020030
  3. Kim J, Choi KB, Park JH, Kim KH. Metabolite profile changes and increased antioxidative and antiinflammatory activities of mixed vegetables after fermentation by Lactobacillus plantarum. PLoS One. 2019; 14(5):e0217180. https://doi.org/10.1371/journal.pone.0217180
  4. Yonezawa S, Xiao JZ, Odamaki T, Ishida T, Miyaji K, Yamada A, Yaeshima T, Iwatsuki K. Improved growth of bifidobacteria by cocultivation with Lactococcus lactis subspecies lactis. J Dairy Sci. 2010; 93(5):1815–1823. https://doi.org/10.3168/jds.2009-2708
  5. Garmasheva I, Vasyliuk O, Kovalenko N, Oleschenko L. New approach for fast screening of lactic acid bacteria for vegetable fermentation. J Microbiol Biotech Food Sci. 2019; 8(4):1066–1071. https://doi.org/10.15414/jmbfs.2019.8.4.1066-1071
  6. Vasyliuk OM, Garmasheva IL, Kovalenko NK. Probiotic properties of strains Lactobacillus plantarum isolated from fermented products. Microbiol & Biotechnol. 2014; 3:23–30.
  7. De Man JD, Rogosa M, Sharpe ME. Medium for the cultivation of lactobacilli. J Appl Bacteriol. 1960; 23(1):130–135. https://doi.org/10.1111/j.1365-2672.1960.tb00188.x
  8. Vasyliuk OM, Kovalenko NK, Harmasheva IL. [Antagonistic properties of Lactobacillus plantarum strains, isolated from traditional fermented products of Ukraine]. Mikrobiol Z. 2014; 76(3):24–30. Ukrainian.
  9. Guevarra RB, Barraquio VL. Viable Counts of Lactic Acid Bacteria In: Philippine Commercial Yogurts. Int J Dairy Sci Process. 2015; 2(5):24–28. https://doi.org/10.19070/2379-1578-150008
  10. Filannino P, Cardinali G, Rizzello CG, Buchin S, De Angelis M, Gobbetti M, Di Cagno R. Metabolic Responses of Lactobacillus plantarum Strains during Fermentation and Storage of Vegetable and Fruit Juices. Appl Environ Microbiol. 2014; 80(7):2206–2215. https://doi.org/10.1128/AEM.03885-13
  11. Plumed-Ferrer C, Koistinen KM, Tolonen TL, Lehesranta SJ, Kärenlampi SO, Mäkimattila E, Joutsjoki V, Virtanen V, von Wright A. Comparative Study of Sugar Fermentation and Protein Expression Patterns of Two Lactobacillus plantarum Strains Grown in Three Different Media. Appl Environm Microbiol. 2008; 74(17):5349–5358. https://doi.org/10.1128/AEM.00324-08
  12. Klaver FAM, Kingma F, Weerkamp AH. Growth and survival of bifidobacteria in milk. Neth Milk Dairy J. 1993; 47:151–164.
  13. Di Cagno R, De Angelis M, Calasso M, Vincentini O, Vernocchi P, Ndagijimana M, De Vincenzi M, Dessı MR, Guerzoni ME, Gobbetti M. Quorum sensing in sourdough Lactobacillus plantarum DC400: induction of plantaricin A (PlnA) under co-cultivation with other lactic acid bacteria and effect of PlnA on bacterial and Caco-2 cells. Proteomics. 2010; 10(11):2175–2190. https://doi.org/10.1002/pmic.200900565
  14. Rojo-Bezares B, Sáenz Y, Navarro L, Zarazaga M, Ruiz-Larrea F, Torres C. Coculture-inducible bacteriocin activity of Lactobacillus plantarum strain J23 isolated from grape mut. Food Microbiol. 2007; 24(5):482–491. https://doi.org/10.1016/j.fm.2006.09.003
  15. Maldonado-Barragán A, Caballero-Guerrero B, Lucena-Padrós H, Ruiz-Barba JL. Induction of bacteriocin production by coculture is widespread among plantaricin-producing Lactobacillus plantarum strains with different regulatory operons. Food Microbiology. 2013; 33(1):40–47. https://doi.org/10.1016/j.fm.2012.08.009
  16. Tabasco R, García-Cayuela T, Peláez C, Requena T. Lactobacillus acidophilus La-5 increases lactacin B production when it senses live target bacteria. Int J Food Microbiol. 2009; 132(2–3):109–116. https://doi.org/10.1016/j.ijfoodmicro.2009.04.004
  17. Svetoch EA, Eruslanov BV, Perelygin VV, Levchuk VP, Seal BS, Stern NJ. Inducer bacteria, unique signal peptides, and low-nutrient media stimulate in vitro bacteriocin production by Lactobacillus spp. and Enterococcus spp. strains. J Agricult Food Chem. 2010; 58(10):6033–6038. https://doi.org/10.1021/jf902802z
  18. Guerra N, Pastrana L. Enhanced Nisin and Pediocin production on whey supplemented with different nitrogen sources. Biotechnol Lett. 2001; 23:609–612. https://doi.org/10.1023/A:1010324910806
  19. Verluyten J, Messens W, De Vuyst L. Sodium chloride reduces production of curvacin A, a bacteriocin produced by Lactobacillus curvatus strain LTH 1174, originating from fermented sausage. Appl Environ Microbiol. 2004a; 70(4):2271–2278. https://doi.org/10.1128/AEM.70.4.2271-2278.2004
  20. Jiménez-Díaz R, Rios-Sánchez RM, Desmazeaud M, Ruiz-Barba JL, Piard JC. Plantaricin S and T, two new bacteriocins produced by Lactobacillus plantarum LPCO10 isolated from a green olive fermentation. Appl Environ Microbiol. 1993; 59(5):1416–1424. https://doi.org/10.1128/AEM.59.5.1416-1424.1993
  21. Leal-Sánchez MV, Jiménez-Diaz R, Maldonado-Barragán A, Garrido-Fernández A, Ruiz-Barba JL. Optimization of bacteriocin production by batch fermentation of Lactobacillus plantarum LPCO10. Appl Environ Microbiol. 2002; 68(9):4465–4471. https://doi.org/10.1128/AEM.68.9.4465-4471.2002
  22. Calasso M, Di Cagno R, De Angelis M, Campanella D, Minervini F, Gobbetti M. Effects of the Peptide Pheromone Plantaricin A and Cocultivation with Lactobacillus sanfranciscensis DPPMA174 on the Exoproteome and the Adhesion Capacity of Lactobacillus plantarum DC400. Appl Environ Microbiol. 2013; 79(8):2657–2669. https://doi.org/10.1128/AEM.03625-12
  23. Henning C, Gautam D, Muriana P. Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array. Microorganisms. 2015; 3(1):1–16. https://doi.org/10.3390/microorganisms3010001
  24. Ruiz-Barba JL, Caballero-Guerrero B, Maldonado-Barragán A, Jiménez-Díaz R. Coculture with specific bacteria enhances survival of Lactobacillus plantarum NC8, an autoinducer-regulated bacteriocin producer, in olive fermentations. Food Microbiol. 2010; 27(3):413–417. https://doi.org/10.1016/j.fm.2009.10.002