World Journal of Microbiology and Biotechnology

, Volume 27, Issue 2, pp 371–379

Characterization and electrotransformation of Lactobacillus plantarum and Lactobacillus paraplantarum isolated from fermented vegetables

Original Paper


Aims of the study were to characterize two Lactobacillus plantarum-related strains, Lact.plantarum and Lactobacillus paraplantarum isolated from fermented vegetables and, for their potential use as starter strains, compare their growth in various food matrices. Species-level identification of the strains belonging to the Lact. plantarum group was performed by multiplex-PCR with species-specific primers and generation of distinct genotypic profiles was carried out by PFGE-based DNA-fingerprinting. Growth profiles were determined in various food and feed matrices. Compared to Lact. plantarum,Lact. paraplantarum reached higher cell densities in all plant-based matrices and MRS broth. On the contrary to the good growth in plant-based matrices and MRS, poor growth was observed in unprocessed milk. Supplemented lactose did not improve the growth of either tested strain, while predigestion of milk proteins with Lactobacillus helveticus or addition of casitone proved to be an effective means to enhance growth. To find out the applicability of molecular methods, the strains were transformed with replicative plasmids by electroporation. To our knowledge, this is a first report of the electrotransformation of Lact. paraplantarum with a recombinant plasmid.


Vegetable fermentation Starter culture Characterization Lactobacillus plantarum Lactobacillus paraplantarum Electrotransformation 


  1. Bates EE, Gilbert HJ, Hazlewood GP, Huckle J, Laurie JI, Mann SP (1989) Expression of a Clostridium thermocellum endoglucanase gene in Lactobacillus plantarum. Appl Environ Microbiol 55:2095–2097Google Scholar
  2. Bringel F, Hubert J-C (1990) Optimized transformation by electroporation of Lactobacillus plantarum strains with plasmid vectors. Appl Microbiol Biotechnol 33:664–670CrossRefGoogle Scholar
  3. Bringel F, Curk M-C, Hubert J-C (1996) Characterization of lactobacilli by Southern-type hybridization with a Lactobacillus plantarum pyrDFE probe. Int J Syst Bacteriol 46:588–594CrossRefGoogle Scholar
  4. Cogan TM, Beresford TP (2002) Microbiology of hard cheese. In: Robinson RK (ed) Dairy microbiology handbook: the microbiology of milk and milk products, 3rd edn. Wiley, New York, pp 515–560Google Scholar
  5. Curk M-C, Hubert J-C, Bringel F (1996) Lactobacillus paraplantarum sp. nov., a new species related to Lactobacillus plantarum. Int J Syst Bacteriol 46:595–598CrossRefGoogle Scholar
  6. Desai P, Sheth T (1997) Controlled fermentation of vegetables using mixed inoculum of lactic cultures. J Food Sci Technol 34:155–158Google Scholar
  7. Fleming HP (1991) Mixed cultures in vegetable fermentations. In: Zeikus JG, Johnson EA (eds) Mixed cultures in biotechnogy. McGraw-Hill, New York, pp 69–102Google Scholar
  8. Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580CrossRefGoogle Scholar
  9. Herias MV, Hessle C, Telemo E, Midtvedt T, Hanson LA, Wold AE (1999) Immunomodulatory effects of Lactobacillus plantarum colonizing the intestine of gnotobiotic rats. Clin Exp Immunol 116:283–290CrossRefGoogle Scholar
  10. Kahala M, Palva A (1999) The expression signals of the Lactobacillus brevis slpA gene direct efficient heterologous protein production in lactic acid bacteria. Appl Microbiol Biotechnol 51:71–78CrossRefGoogle Scholar
  11. Karovicová J, Drdák M, Greif G, Hybenová E (1999) The choice of strains of Lactobacillus species for the lactic acid fermentation of vegetable juices. Eur Food Res Technol 210:53–56CrossRefGoogle Scholar
  12. Khalid NM, Marth EH (1990) Purification and partial characterization of a prolyl-dipeptidyl aminopeptidase from Lactobacillus helveticus CNRZ 32. Appl Environ Microbiol 56:381–388Google Scholar
  13. Kleerebezem M, Boekhorst J, van Kranenburg R, Molenaar D, Kuipers OP, Leer R, Tarchini R, Peters SA, Sandbrink HM, Fiers MW, Stiekema W, Lankhorst RM, Bron PA, Hoffer SM, Groot MN, Kerkhoven R, de Vries M, Ursing B, de Vos WM, Siezen RJ (2003) Complete genome sequence of Lactobacillus plantarum WCFS1. Proc Natl Acad Sci USA 100:1990–1995CrossRefGoogle Scholar
  14. Koistinen KM, Plumed-Ferrer C, Lehesranta SJ, Kärenlampi SO, von Wright A (2007) Comparison of growth-phase-dependent cytosolic proteomes of two Lactobacillus plantarum strains used in food and feed fermentations. FEMS Microbiol Lett 273:12–21CrossRefGoogle Scholar
  15. Kok J, van der Vossen JM, Venema G (1984) Construction of plasmid cloning vectors for lactic streptococci which also replicate in Bacillus subtilis and Escherichia coli. Appl Environ Microbiol 48:726–731Google Scholar
  16. Leal MV, Baras M, Ruiz-Barba JL, Floriano B, Jiménez-Diáz R (1998) Bacteriocin production and competitiveness of Lactobacillus plantarum LPCO10 in olive juice broth, a culture medium obtained from olives. Int J Food Microbiol 43:129–134CrossRefGoogle Scholar
  17. Mäki M (2004) Lactic acid bacteria in vegetable fermentations. In: Salminen S, von Wright A, Ouwehand A (eds) Lactic acid bacteria: microbiological and functional aspects, 2nd edn. Marcel Dekker, Inc., New York, pp 419–430. Revised and ExpandedGoogle Scholar
  18. McDonald LC, Fleming HP, Hassan HM (1990) Acid tolerance of Leuconostoc mesenteroides and Lactobacillus plantarum. Appl Environ Microbiol 56:2120–2124Google Scholar
  19. Park WJ, Lee KH, Lee JM, Lee HJ, Kim JH, Lee J-H, Chang HC, Chung DK (2004) Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi and development of lactic acid bacteria-Escherichia coli shuttle vector. Plasmid 52:84–88CrossRefGoogle Scholar
  20. Pavan S, Hols P, Delcour J, Geoffroy M-C, Grangette C, Kleerebezem M, Mercenier A (2000) Adaptation of the nisin-controlled expression system in Lactobacillus plantarum: a tool to study in vivo biological effects. Appl Environ Microbiol 66:4427–4432CrossRefGoogle Scholar
  21. Plumed-Ferrer C, Kivelä I, Hyvönen P, von Wright A (2005) Survival, growth and persistence under farm conditions of a Lactobacillus plantarum strain inoculated into liquid pig feed. J Appl Microbiol 99:851–858CrossRefGoogle Scholar
  22. Posno M, Leer RJ, van Luijk N, van Giezen MJ, Heuvelmans PT, Lokman BC, Pouwels PH (1991) Incompatibility of Lactobacillus vectors with replicons derived from small cryptic Lactobacillus plasmids and segregational instability of the introduced vectors. Appl Environ Microbiol 57:1822–1828Google Scholar
  23. Rose AH (1982) Economic microbiology, fermented foods. Academic Press, LondonGoogle Scholar
  24. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  25. Scheirlinck T, Mahillon J, Joos H, Dhaese P, Michiels F (1989) Integration and expression of α-amylase and endoglucanase genes in the Lactobacillus plantarum chromosome. Appl Environ Microbiol 55:2130–2137Google Scholar
  26. Scheirlinck T, De Meutter J, Arnaut G, Joos H, Claeyssens M, Michiels F (1990) Cloning and expression of cellulase and xylanase genes in Lactobacillus plantarum. Appl Microbiol Biotechnol 33:534–541CrossRefGoogle Scholar
  27. Tamminen M, Mäki M, Joutsjoki T, Ryhänen E-L, Joutsjoki V (2003) Differentiation of lactobacilli related to Lactobacillus plantarum from naturally fermented cucumbers and white cabbage. Appl Biotechnol Food Sci Policy 1:125–128Google Scholar
  28. Tamminen M, Joutsjoki T, Sjöblom M, Joutsen M, Palva A, Ryhänen E-L, Joutsjoki V (2004) Screening of lactic acid bacteria from fermented vegetables by carbohydrate profiling and PCR-ELISA. Lett Appl Microbiol 39:439–444CrossRefGoogle Scholar
  29. 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 67:3450–3454CrossRefGoogle Scholar
  30. Tynkkynen S, Satokari R, Saarela M, Mattila-Sandholm T, Saxelin M (1999) Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains. Appl Environ Microbiol 65:3908–3914Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Elina Mäkimattila
    • 1
  • Minna Kahala
    • 1
  • Vesa Joutsjoki
    • 1
  1. 1.MTT Agrifood Research Finland, Biotechnology and Food ResearchJokioinenFinland

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