Skip to main content
SpringerLink
Log in

Functionality of the S-layer protein from the probiotic strain Lactobacillus helveticus M92

  • Original Paper
  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

The objective of this study was the characterisation of the S-layer protein (SlpA) and its functional role in the probiotic activity of Lactobacillus helveticus M92. SlpA was isolated and identified by SDS-PAGE LC-MS/MS analysis. The slpA gene encoding the SlpA from L. helveticus M92 was sequenced and compared with other well characterised slpA genes. Sequence similarity searches revealed high homology with the SlpA of Lactobacillus strains. Purified SlpA showed significantly better immunomodulatory effects in orally immunised mice than L. helveticus M92 cells after SlpA removal. SlpA is involved in the autoaggregation of L. helveticus M92 cells and coaggregation of L. helveticus M92 with S. Typhimurium FP1 as these processes were negatively affected after SlpA removal from the cell surface. Therefore, the influence of oral treatment with L. helveticus M92 on an oral infection of mice by S. Typhimurium FP1 was investigated. Following the oral immunization of mice, with viable L. helveticus M92 and S. Typhimurium FP1 cells, the concentration in the luminal contents of total S-IgA and specific anti-Salmonella S-IgA antibodies, from all immunized mice was significantly higher compared to the control group or a group of mice infected only with S. Typhimurium FP1. These results demonstrate that the observed reduced infection by S. Typhimurium FP1 in mice with L. helveticus M92 is associated with competitive exclusion in the intestinal tract and enhanced immune protection conferred by the L. helveticus M92 and its SlpA.

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

Access this article

Log in via an institution

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
Fig. 5

Similar content being viewed by others

References

  • Åvall-Jääskeläinen S, Palva A (2005) Lactobacillus surface layers and their applications. FEMS Microbiol Rev 29:511–529

    Article  PubMed  Google Scholar 

  • Åvall-Jääskeläinen S, Hynönen U, Ilk N, Pum D, Sleytr UB, Palva A (2008) Identification and characterization of domains responsible for self-assembly and cell wall binding of the surface layer protein of Lactobacillus brevis ATCC 8287. BMC Microbiol 8(165):1–15

    Google Scholar 

  • Beganović J (2008) Application of proteomics and other molecular methods in the characterisation of functionality of the probiotic bacteria. Dissertation, University of Zagreb

  • Beganović J, Guillot A, van de Guchte M, Jouan A, Gitton C, Loux V, Roy K, Huet S, Monod H, Monnet V (2010) Characterization of the insoluble proteome of Lactococcus lactis by SDS-PAGE LC-MS/MS leads to the identification of new markers of adaptation of the bacteria to the mouse digestive tract. J Proteome Res 9:677–688

    Article  PubMed  Google Scholar 

  • Boesten RJ, de Vos WM (2008) Interactomics in the human intestine: Lactobacilli and Bifidobacteria make a difference. J Clin Gastroenterol 42(Suppl 3 Pt 2):S163–S167

    Article  PubMed  Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Buck BL, Altermann E, Svingerud T, Klaenhammer TR (2005) Functional analysis of putative adhesion factors in Lactobacillus acidophilus NCFM. Appl Environ Microbiol 71(12):8344–8351

    Article  PubMed  CAS  Google Scholar 

  • Casey PG, Casey GD, Gardiner GE, Tangney M, Stanton C, Ross RP, Hill C, Fitzgerald GF (2004) Isolation and characterisation of anti-Salmonella lactic acid bacteria from porcine gastrointestinal tract. Lett Appl Microbiol 39:431–438

    Article  PubMed  CAS  Google Scholar 

  • Chen XY, Xu JJ, Shuai JB, Chen JS, Zhang ZF, Fang WH (2007) The S-layer proteins of Lactobacillus crispatus strain ZJ001 is responsible for competitive exclusion against Escherichia coli O157: H7 and Salmonella typhimurium. Int J Food Microbiol 115:307–312

    Article  PubMed  CAS  Google Scholar 

  • Coconnier MH, Lievin V, Lorrot M, Servin AL (2000) Antagonistic activity of Lactobacillus acidophilus LB against intracellular Salmonella enterica serovar Typhimurium infecting human enterocyte-like Caco-2/TC-7 cells. Appl Environ Microbiol 66:1152–1157

    Article  PubMed  CAS  Google Scholar 

  • Delcour J, Ferain T, Deghorain M, Palumbo E, Hols P (1999) The biosynthesis and functionality of the cell-wall of lactic acid bacteria. Antonie van Leeuwenhoek 76:159–184

    Article  PubMed  CAS  Google Scholar 

  • Fayol-Messaoudi D, Berger CN, Coconnier-Polter MH, Liévin-Le Moal V, Servin AL (2005) pH-, lactic acid-, and non-lactic acid-dependent activities of probiotic lactobacilli against Salmonella enterica Serovar Typhimurium. Appl Environ Microbiol 71(10):6008–6013

    Article  PubMed  CAS  Google Scholar 

  • Frece J, Kos B, Svetec IK, Zgaga Z, Mrsa V, Šušković J (2005a) Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92. J Appl Microbiol 98:285–292

    Article  PubMed  CAS  Google Scholar 

  • Frece J, Kos B, Beganović J, Vuković S, Šušković J (2005b) In vivo testing of functional properties of three selected probiotic strains. World J Microbiol Biotechnol 21:1401–1408

    Article  Google Scholar 

  • Frece J, Kos B, Svetec IK, Zgaga Z, Beganović J, Leboš A, Šušković J (2009) Synbiotic effect of Lactobacillus helveticus M92 and prebiotics on the intestinal microflora and immune system of mice. J Dairy Res 76:98–104

    Article  PubMed  CAS  Google Scholar 

  • Golowczyc MA, Mobili P, Garrote GL, Abraham AG, De Antoni GL (2007) Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteridis. Int J Food Microbiol 118:264–273

    Article  PubMed  CAS  Google Scholar 

  • Grogono-Thomas R, Blaser MJ, Ahmadi M, Newell DG (2003) Role of S-layer protein antigenic diversity in the immune responses of sheep experimentally challenged with Campylobacter fetus subsp. Fetus. Infect Immun 71:147–154

    Article  PubMed  CAS  Google Scholar 

  • Hajduk G, Kos B, Šušković J, Frece J, Leboš A, Beganović J (2009) Probiotic properties of Bifidobacterium animalis subsp. lactis BB-12 in baby cereal flakes enriched with inulin. Ital J Food Sci 4(21):473–486

    Google Scholar 

  • Horie M, Kajikawa HS, Toba T (2002) Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene. Lett Appl Microbiol 35(1):57–61

    Article  PubMed  CAS  Google Scholar 

  • Johnson-Henry KC, Hagen KE, Gordonpour M, Tompkins TA, Sherman PM (2007) Surface-layer protein extracts from Lactobacillus helveticus inhibit enterohaemorrhagic Escherichia coli O157:H7 adhesion to epithelial cells. Cell Microbiol 9(2):356–367

    Article  PubMed  CAS  Google Scholar 

  • Kos B, Šušković J, Goreta J, Matošić S (2000) Effect of protectors on the viability of Lactobacillus acidophilus M92 in simulated gastrointestinal conditions. Food Technol Biotechnol 36:121–127

    Google Scholar 

  • Kos B, Šušković J, Vuković S, Šimpraga M, Frece J, Matošić S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987

    Article  PubMed  CAS  Google Scholar 

  • Kos B, Šušković J, Beganović J, Gjuračić K, Frece J, Iannaccone C, Canganella F (2008) Characterization of the three selected probiotic strains for the application in food industry. World J Microbiol Biotechnol 24:699–707

    Article  CAS  Google Scholar 

  • Lebeer S, Vanderleyden J, De Keersmaecker SCJ (2008) Genes and molecules of lactobacilli supporting probiotic action. Microbiol Mol Biol Rev 72(4):728–764

    Article  PubMed  CAS  Google Scholar 

  • Leboš Pavunc A, Beganović J, Kos B, Buneta A, Beluhan S, Šušković J (2010) Influence of microencapsulation and transglutaminase on viability of probiotic strain Lactobacillus helveticus M92 and consistency of set yoghurt. Int J Dairy Techol 76 (in press). doi:10.1111/j.1471-0307.2010.00647.x

  • Liu TS, P Zhang, Ma Y, Qin H (2010) Lactobacillus plantarum surface layer adhesive protein protects intestinal epithelial cells against tight junction injury induced by enteropathogenic Escherichia coli. Mol Biol Rep. doi:10.1007/s11033-010-0457-8

  • Martínez B, Sillanpää J, Smit E, Korhonen TK, Pouwels PH (2000) Expression of cbsA encoding the collagen-binding S-protein of Lactobacillus crispatus JCM5810 in Lactobacillus casei ATCC 393T. J Bacteriol 182(23):6857–6861

    Article  PubMed  Google Scholar 

  • Mobili P, Serradell MA, Trejo SA, Puigvert FXA, Abraham GA GL, Antoni De (2009) Heterogeneity of S-layer proteins from aggregating and non-aggregating Lactobacillus kefir strains. Antonie van Leeuwenhoek 95:363–372

    Article  PubMed  CAS  Google Scholar 

  • National Research Council (1996) Guide for the care and use of laboratory animals. Institute of Laboratory Animal Resources, National Academy Press, Washington, DC

  • Perdigón G, Locascio M, Medici M, Pesce de Ruiz Holgado A, Oliver G (2003) Interaction of bifidobacteria with the gut and their influence in the immune function. Biocell 27:1–9

    PubMed  Google Scholar 

  • Racedo S, Villena J, Medina M, Agüero G, Rodríguez V, Alvarez S (2006) Lactobacillus casei administration reduces lung injuries in a Streptococcus pneumoniae infection in mice. Microbes Infect 8:2359–2366

    Article  PubMed  Google Scholar 

  • Servin AL, Coconnier MH (2003) Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens. Best Pract Res Clin Gastroenterol 17(5):741–754

    Article  PubMed  CAS  Google Scholar 

  • Šušković J, Kos B, Matošić S, Besendorfer V (2000) The effect of bile salts on survival and morphology of potential probiotic strain Lactobacillus acidophilus M92. World J Microbiol Biotechnol 16:673–678

    Article  Google Scholar 

  • Šušković J, Kos B, Goreta J, Matošić S (2001) Role of lactic acid bacteria and bifidobacteria in synbiotic effect. Food Technol Biotechnol 39:227–235

    Google Scholar 

  • Šušković J, Kos B, Beganović J, Leboš Pavunc A, Habjanič K, Matošić S (2010) Antimicrobial activity—the most important property of probiotic and starter lactic acid bacteria. Food Technol Biotechnol 48(3):296–307

    Google Scholar 

  • Van der Mei HC, van de Belt-Gritter B, Pouwels PH, Martinez B, Busscher HJ (2003) Cell surface hydrophobicity is conveyed by S-layer: a study in recombinant lactobacilli. Colloids Surf B Biointerfaces 28:127–134

    Article  Google Scholar 

  • Vidgrén G, Palva I, Pakkanen R, Lounatmaa K, Palva A (1992) S-layer protein gene of Lactobacillus brevis: cloning by polymerase chain reaction and determination of the nucleotide sequence. J Bacteriol 174(22):7419–7427

    PubMed  Google Scholar 

  • Yoneda M, Hirofuji T, Motooka N, Nozoe K, Shigenaga K, Anan H, Miura M, Kabashima H, Matsumoto A, Maeda K (2003) Humoral immune responses to S-layer-like proteins of Bacteroides forsythus. Clin Diagn Lab Immunol 10:383–387

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful for the financial support provided by Ministry of Science, Education and Sports of the Republic of Croatia (Project 0581990 “Probiotics, prebiotics and functional starter cultures”). The authors wish also to thank to the staff of PAAPSO, INRA Jouy en Josas, France for mass spectrometry analysis. Jasna Beganović was recipient of a Marie Curie fellowship for Early Stage Research Training, inside LABHEALTH (MEST-CT-2004-514428).

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Department of Biochemical Engineering, Laboratory for Antibiotic, Enzyme, Probiotic and Starter Culture Technology, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000, Zagreb, Croatia

    Jasna Beganović, Jadranka Frece, Blaženka Kos, Andreja Leboš Pavunc, Ksenija Habjanič & Jagoda Šušković

Authors
  1. Jasna Beganović
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jadranka Frece
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Blaženka Kos
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andreja Leboš Pavunc
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ksenija Habjanič
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jagoda Šušković
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Blaženka Kos.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Beganović, J., Frece, J., Kos, B. et al. Functionality of the S-layer protein from the probiotic strain Lactobacillus helveticus M92. Antonie van Leeuwenhoek 100, 43–53 (2011). https://doi.org/10.1007/s10482-011-9563-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10482-011-9563-4

Keywords

Search

Navigation