Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral streptococci biofilm


Lactic acid bacteria (LAB) can interfere with pathogens through different mechanisms; one is the production of biosurfactants, a group of surface-active molecules, which inhibit the growth of potential pathogens. In the present study, biosurfactants produced by Lactobacillus reuteri DSM 17938, Lactobacillus acidophilus DDS-1, Lactobacillus rhamnosus ATCC 53103, and Lactobacillus paracasei B21060 were dialyzed (1 and 6 kDa) and characterized in term of reduction of surface tension and emulsifying activity. Then, aliquots of the different dialyzed biosurfactants were added to Streptococcus mutans ATCC 25175 and Streptococcus oralis ATCC 9811 in the culture medium during the formation of biofilm on titanium surface and the efficacy was determined by agar plate count, biomass analyses, and flow cytometry. Dialyzed biosurfactants showed abilities to reduce surface tension and to emulsifying paraffin oil. Moreover, they significantly inhibited the adhesion and biofilm formation on titanium surface of S. mutans and S. oralis in a dose-dependent way, as demonstrated by the remarkable decrease of cfu/ml values and biomass production. The antimicrobial properties observed for dialyzed biosurfactants produced by the tested lactobacilli opens future prospects for their use against microorganisms responsible of oral diseases.

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

Fig. 1
Fig. 2
Fig. 3


  1. Ahimou F, Jacques P, Deleu M (2000) Surfactin and iturin a effects on Bacillus subtilis surface hydrophobicity. Enzym Microb Technol 27:749–754. doi:10.1016/S0141-0229(00)00295-7

  2. Banat I, Franzetti A, Gandolfi I, Bestetti G, Martinotti M, Fracchia L, Smyth T, Marchant R (2010) Microbial biosurfactants production, applications and future potential. Appl Microbiol Biotechnol 87:427–444. doi:10.1007/s00253-010-2589-0

  3. Banat IM, Makkar RS, Cameotra SS (2000) Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol 53:495–508

  4. Barbesti S, Citterio S, Labra M, Baroni MD, Neri MG, Sgorbati S (2000) Two and three-color fluorescence flow cytometric analysis of immunoidentified viable bacteria. Cytometry 40:214–218. doi:10.1002/1097-0320(20000701)40:3<214::AID-CYTO6>3.0.CO;2-M

  5. Çaglar E, Cildir SK, Ergeneli S, Sandalli N, Twetman S (2006) Salivary mutans streptococci and lactobacilli levels after ingestion of the probiotic bacterium Lactobacillus reuteri ATCC 55730 by straws or tablets. Acta Odontol Scand 64:314–318. doi:10.1080/00016350600801709

  6. Çaglar E, Kuscu OO, Cildir SK, Kuvvetli SS, Sandalli N (2008) A probiotic lozenge administered medical device and its effect on salivary mutans streptococci and lactobacilli. Int J Paediatr Dent 18:35–39. doi:10.1111/j.1365-263X.2007.00866.x

  7. Ciandrini E, Campana R, Federici S, Manti A, Battistelli M, Falcieri E, Papa S, Baffone W (2014) In vitro activity of Carvacrol against titanium-adherent oral biofilms and planktonic cultures. Clin Oral Investig 18:2001–2013. doi:10.1007/s00784-013-1179-9

  8. Daverey A, Pakshirajan K (2009) Production, characterization, and properties of sophorolipids from the yeast Candida bombicola using a low-cost fermentative medium. Appl Biochem Biotechnol 158:663–674. doi:10.1007/s12010-008-8449-z

  9. Elter C, Heuer W, Demling A, Hannig M, Heidenblut T, Bach F-W, Stiesch-Scholz M (2008) Supra- and subgingival biofilm formation on implant abutments with different surface characteristics. Int J Oral Maxillofac Implants 23:327–334

  10. Francy DS, Thomas JM, Raymond RL, Ward CH (1991) Emulsification of hydrocarbons by subsurface bacteria. J Ind Microbiol 8:237–245. doi:10.1007/BF01576061

  11. Fürst MM, Salvi GE, Lang NP, Persson GR (2007) Bacterial colonization immediately after installation on oral titanium implants. Clin Oral Implants Res 18:501–508. doi:10.1111/j.1600-0501.2007.01381.x

  12. Gomaa EZ (2013) Antimicrobial and anti-adhesive properties of biosurfactant produced by lactobacilli isolates, biofilm formation and aggregation ability. J Gen Appl Microbiol 59:425–436. doi:10.2323/jgam.59.425

  13. Gudiña EJ, Teixeira JA, Rodrigues LR (2010) Isolation and functional characterization of a biosurfactant produced by Lactobacillus paracasei. Colloids Surf B Biointerfaces 76:298–304. doi:10.1016/j.colsurfb.2009.11.008

  14. Haukioja A, Loimaranta V, Tenovuo J (2008) Probiotic bacteria affect the composition of salivary pellicle and streptococcal adhesion in vitro. Oral Microbiol Immunol 23:336–343. doi:10.1111/j.1399-302X.2008.00435.x

  15. Jalasvuori H, Haukioja A, Tenovuo J (2012) Probiotic Lactobacillus reuteri strains ATCC PTA 5289 and ATCC 55730 differ in their cariogenic properties in vitro. Arch Oral Biol 57:1633–1638. doi:10.1016/j.archoralbio.2012.07.014

  16. Jeon J-G, Rosalen PL, Falsetta ML, Koo H (2011) Natural products in caries research: current (limited) knowledge, challenges and future perspective. Caries Res 45:243–263

  17. Kim SH, Lim EJ, Lee SO, Lee JD, Lee TH (2000) Purification and characterization of biosurfactants from Nocardia sp. L-417. Biotechnol Appl Biochem 31:249–253

  18. Kuiper I, Lagendijk EL, Pickford R, Derrick JP, Lamers GEM, Thomas-Oates JE, Lugtenberg BJJ, Bloemberg GV (2004) Characterization of two Pseudomonas putida lipopeptide biosurfactants, putisolvin I and II, which inhibit biofilm formation and break down existing biofilms. Mol Microbiol 51:97–113. doi:10.1046/j.1365-2958.2003.03751.x

  19. Marttinen A, Haukioja A, Keskin M, Söderling E (2013) Effects of Lactobacillus reuteri PTA 5289 and L. paracasei DSMZ16671 on the adhesion and biofilm formation of Streptococcus mutans. Curr Microbiol 67:193–199. doi:10.1007/s00284-013-0352-3

  20. Meurman JH, Stamatova I (2011) Lactic acid bacteria in oral health. in: lactic acid bacteria in oral health, lactic acid bacteria: microbiological and functional aspects. CRC Press, pp 403–422

  21. Myers D (2005) Surfactant science and technology, 3rd edn. Wiley, Newyork

  22. Nitschke M, Costa SGVAO, Contiero J (2005) Rhamnolipid surfactants: an update on the general aspects of these remarkable biomolecules. Biotechnol Prog 21:1593–1600. doi:10.1021/bp050239p

  23. Ntrouka VI, Slot DE, Louropoulou A, Van der Weijden F (2011) The effect of chemotherapeutic agents on contaminated titanium surfaces: a systematic review. Clin Oral Implants Res 22:681–690. doi:10.1111/j.1600-0501.2010.02037.x

  24. De la Fuente-Núñez C, Reffuveille F, Fernández L, Hancock REW (2013) Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 16:580–589. doi:10.1016/j.mib.2013.06.013

  25. Rodrigues L, Banat IM, Teixeira J, Oliveira R (2006a) Biosurfactants: potential applications in medicine. J Antimicrob Chemother 57:609–618. doi:10.1093/jac/dkl024

  26. Rodrigues LR, Teixeira JA, van der Mei HC, Oliveira R (2006b) Isolation and partial characterization of a biosurfactant produced by Streptococcus thermophilus a. Colloids Surf B Biointerfaces 53:105–112. doi:10.1016/j.colsurfb.2006.08.009

  27. Saharan BS, Sahu RK, Sharma D (2012) A review on biosurfactants: fermentation, current developments and perspectives. Genet Eng Biotechnol J 1:1–14

  28. Saravanakumari P, Mani K (2010) Structural characterization of a novel xylolipid biosurfactant from Lactococcus lactis and analysis of antibacterial activity against multi-drug resistant pathogens. Bioresour Technol 101:8851–8854. doi:10.1016/j.biortech.2010.06.104

  29. Sharma D, Saharan BS (2014) Simultaneous production of biosurfactants and bacteriocins by probiotic Lactobacillus casei MRTL3. Int J Microbiol Article ID 698713 . doi:10.1155/2014/6987137 pages

  30. Sharma D, Saharan B, Chauhan N, Procha S, Lal S (2015) Isolation and functional characterization of novel biosurfactant produced by Enterococcus faecium. SpringerPlus 4:4

  31. Sharma D, Saharan BS, Chauhan N, Bansal A, Procha S (2014) Production and structural characterization of Lactobacillus helveticus derived biosurfactant. Sci World J Article ID:493548 . doi:10.1155/2014/4935489 pages

  32. Socransky SS, AD H (2002) Dental biofilms: difficult therapeutic targets. Periodontol 2000 28:12–55

  33. Söderling E, Marttinen A, Haukioja A (2011) Probiotic lactobacilli interfere with Streptococcus mutans biofilm formation in vitro. Curr Microbiol 62:618–622. doi:10.1007/s00284-010-9752-9

  34. Spinler JK, Taweechotipatr M, Rognerud CL, Ou CN, Tumwasorn S, Versalovic J (2008) Human-derived probiotic Lactobacillus reuteri demonstrate antimicrobial activities targeting diverse enteric bacterial pathogens. Anaerobe 14:166–171. doi:10.1016/j.anaerobe.2008.02.001

  35. Tahmourespour A, Salehi R, Kermanshahi RK (2011) Lactobacillus acidophilus-derived biosurfactant effect on gtfb and gtfc expression level in Streptococcus mutans biofilm cells. Braz J Microbiol 42:330–339

  36. Teanpaisan R, Piwat S, Dahlén G (2011) Inhibitory effect of oral Lactobacillus against oral pathogens. Lett Appl Microbiol 53:452–459. doi:10.1111/j.1472-765X.2011.03132.x

  37. Walencka E, Różalska S, Sadowska B, Różalska B (2008) The influence of Lactobacillus acidophilus-derived surfactants on staphylococcal adhesion and biofilm formation. Folia Microbiol (Praha) 53:61–66. doi:10.1007/s12223-008-0009-y

Download references


This work was supported by partial grants from PIO SODALIZIO dei PICENI Foundation (Rome, Italy).

Author information

Correspondence to Wally Baffone.

Ethics declarations

Ethical statement

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Eleonora Ciandrini and Raffaella Campana contributed equally to the research.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ciandrini, E., Campana, R., Casettari, L. et al. Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral streptococci biofilm. Appl Microbiol Biotechnol 100, 6767–6777 (2016). https://doi.org/10.1007/s00253-016-7531-7

Download citation


  • Biosurfactants
  • Lactobacillus
  • Chemical characterization
  • Oral streptococci
  • Biofilm formation percentages