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Adhesion of indigenous Lactobacillus plantarum to gut extracellular matrix and its physicochemical characterization

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Abstract

Adhesion to the human intestinal epithelial cell is considered as one of the important selection criteria of lactobacilli for probiotic attributes. Sixteen Lactobacillus plantarum strains from human origins were subjected for adhesion to extracellular matrix (ECM) components, and their physiochemical characterization, incubation time course and effect of different pH on bacterial adhesion in vitro were studied. Four strains showed significant binding to both fibronectin and mucin. After pretreatment with pepsin and trypsin, the bacterial adhesion to ECM reduced to the level of 50 % and with lysozyme significantly decreased by 65–70 %. Treatment with LiCl also strongly inhibited (90 %) the bacterial adhesion to ECM. Tested strains showed highest binding efficacy at time course of 120 and 180 min. Additionally, the binding of Lp91 to ECM was highest at pH 6 (155 ± 2.90 CFU/well). This study proved that surface layer components are proteinaceous in nature, which contributed in adhesion of lactobacillus strains. Further, the study can provide a better platform for introduction of new indigenous probiotic strains having strong adhesion potential for future use.

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References

  • Adlerberth IS, Ahrne ML, Johansson G, Molin LA (1996) A mannose-specific adherence mechanism in Lactobacillus plantarum conferring binding to the human colonic cell line HT-29. Appl Environ Microbiol 62:2244–2251

    PubMed Central  CAS  PubMed  Google Scholar 

  • Ali QS, Farid AJ, Kabeir BM, Zamberi S, Shuhaimi M, Ghazali HM (2009) Adhesion properties of Bifidobacterium pseudocatenulatum G4 and Bifidobacterium Longum BB536 on HT-29 human epithelium cell line at different times and pH. Proc World Acad Sci Eng Tech 37:2070–3740

    Google Scholar 

  • Antikainen J, Anton L, Sillanpaa J, Korhonen TK (2002) Domains in S-layer protein CbsA of Lactobacillus crispatus involved in adherence to collagens, laminin and lipoteichoic acids and self-assembly. Mol Microbiol 46:381–394

    Article  CAS  PubMed  Google Scholar 

  • Balgir PP, Kaur B, Kaur T, Daroch N, Kaur G (2013) In vitro and in vivo survival and colonic adhesion of Pediococcus acidilactici MTCC5101 in human gut. Biomed Res Int. doi:10.1155/2013/583850

    Google Scholar 

  • Bergonzelli GE, Granato D, Pridmore RD, Marvin-Guy LF, Donnicola D, Corthesy-Theulaz IE (2006) GroEL of Lactobacillus johnsonii La1 (NCC533) is cell surface associated: potential role in interactions with the host and the gastric pathogen Helicobacter pylori. Infect Immun 74:425–434

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Boot HJ, Kolen CPAM, van Noort JM, Pouwels PH (1993) S-layer protein of Lactobacillus acidophilus ATCC 4356: purification, expression in Escherichia coli and nucleotide sequence of the corresponding gene. J Bacteriol 175:6089–6096

    PubMed Central  CAS  PubMed  Google Scholar 

  • Botes M, Loos B, Carol A, Reenen V, Dicks LMT (2008) Adhesion of the probiotic strains Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 to Caco-2 cells under conditions simulating the intestinal tract, and in the presence of antibiotics and anti-inflammatory medicaments. Arch Microbiol 190:573–584

    Article  CAS  PubMed  Google Scholar 

  • Caballero-Franco C, Keller K, De Simone C, Chadee K (2007) The VSL#3 probiotic formula induces mucin gene expression and secretion in colonic epithelial cells. Am J Physiol Gastrointest Liver Physiol 292(1):G315–G322

    Article  CAS  PubMed  Google Scholar 

  • Castaldo C, Vastano V, Siciliano RA, Candela M, Vici M, Muscariello L et al (2009) Surface displaced alfa-enolase of Lactobacillus plantarum is a fibronectin binding protein. Microb Cell Factories. doi:10.1186/1475-2859-8-14

  • Crittenden R, Karppinen S, Ojanen S, Tenkanen M, Fagerstrom R, Matto J et al (2002) In vitro fermentation of cereal dietary fibre carbohydrates by probiotic and intestinal bacteria. J Sci Food Agric 82:781–789

    Article  CAS  Google Scholar 

  • De Leeuw E, Li X, Lu WY (2006) Binding characteristics of the Lactobacillus brevis ATCC 8287 surface layer to extracellular matrix proteins. FEMS Microbiol Lett 260:210–215

    Article  PubMed  Google Scholar 

  • Diego MP, Llopis M, Antolín M, de Torres I, Guarner F, Martínez GP, Monedero V (2009) Adhesion properties of Lactobacillus casei strains to resected intestinal fragments and components of the extracellular matrix. Arch Microbiol 191:153–161

    Article  Google Scholar 

  • Duary RK, Batish VK, Grover S (2010) Expression of the atpD gene in probiotic Lactobacillus plantarum strains under in vitro acidic conditions using RT-qPCR. Res Microbiol 161:399–405

    Article  CAS  PubMed  Google Scholar 

  • Duary RK, Rajput YS, Batish VK, Grover S (2011) Assessing the adhesion of putative indigenous probiotic lactobacilli to human colonic epithelial cells. Indian J Med Res 134:664–671

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Duary RK, Batish VK, Grover S (2012a) Relative gene expression of bile salt hydrolase and surface proteins in two putative indigenous Lactobacillus plantarum strains under in vitro gut conditions. Mol Biol Rep 39(3):2541–2552

    Article  CAS  PubMed  Google Scholar 

  • Duary RK, Bhausaheb MA, Batish VK, Grover S (2012b) Anti-inflammatory and immunomodulatory efficacy of indigenous probiotic Lactobacillus plantarum Lp91 in colitis mouse model. Mol Biol Rep 39(4):4765–4775

    Article  CAS  PubMed  Google Scholar 

  • Elli M, Callegari ML, Ferrari S, Bessi E, Cattivelli D, Soldi S, Morelli L, Feuillerat NG, Antoine JM (2006) Survival of yogurt bacteria in the human gut. Appl Environ Microbiol 72(7):5113–5117

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Granato D, Bergonzelli GE, Pridmore RD, Marvin L, Rouvet M, Corthesy-Theulaz IE (2004) Cell surface-associated elongation factor Tu mediates the attachment of Lactobacillus johnsonii NCC533 (La1) to human intestinal cells and mucins. Infect Immun 72:2160–2169

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Greene JD, Klaenhammer TR (1994) Factors involved in adherence of lactobacilli to human Caco-2 cells. Appl Environ Microbiol 60:4487–4494

    PubMed Central  CAS  PubMed  Google Scholar 

  • Grover S, Kumar A, Srivastava AK, Batish VK (2012) Probiotics as functional food ingredients for augmenting human health. Innovation in Healthy and Functional Foods. CRC Press, Taylor & Francis Group, Boca Raton, pp 387–417

    Chapter  Google Scholar 

  • Hardy H, Harris J, Lyon E, Beal J, Foey AD (2013) Probiotics, prebiotics and immunomodulation of gut mucosal defences: homeostasis and immunopathology. Nutrients 5:1869–1912. doi:10.3390/nu5061869

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hidalgo Maria, Martin-Santamaria Sonsoles, Recio Isidra, Sanchez-Moreno Concepcion, de Pascual-Teresa Beatriz, Rimbach Gerald, de Pascual-Teresa Sonia (2012) Potential anti-inflammatory, anti-adhesive, anti/estrogenic, and angiotensin-converting enzyme inhibitory activities of anthocyanins and their gut metabolites. Genes Nutr 7(2):295–306

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hynonen UB, Westerlund W, Palva A, Korhonen TK (2002) Identification by flagellum display of an epithelial cell- and fibronectin-binding function in the SlpA surface protein of Lactobacillus brevis. J Bacteriol 184:3360–3367

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Jankovic I, Ventura M, Meylan V, Rouvet M, Elli M, Zink R (2003) Contribution of aggregating-promoting factor to maintenance of cell shape in Lactobacillus gasseri 4B2. J Bacteriol 185:3288–3296

    Article  PubMed Central  CAS  PubMed  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 4(12):e8099. doi:10.1371/journal.pone.0008099

    Article  PubMed Central  PubMed  Google Scholar 

  • Kumar R, Grover S, Batish VK (2011) Hypocholesterolaemic effect of dietary inclusion of two putative probiotic bile salt hydrolaseproducing Lactobacillus plantarum strains in Sprague–Dawley rats. Br J Nutr 105:561–573

    Article  CAS  PubMed  Google Scholar 

  • Kumar R, Grover S, Batish VK (2012) Bile salt hydrolase (Bsh) activity screening of lactobacilli. In vitro selection of indigenous lactobacillus strains with potential bile salt hydrolysing and cholesterol-lowering ability. Probiot Antimicrob Proteins 4(3):162–172

    Article  CAS  Google Scholar 

  • Letourneau J, Levesque C, Berthiaume F, Jacques M, Mourez M (2011) In vitro assay of bacterial adhesion onto mammalian epithelial cells. J Vis Exp d16;(51). doi:10.3791/2783

  • Lorca G, Torino MI, de Valdez GF, Ljung A (2002) Lactobacilli express cell surface proteins which mediate binding of immobilized collagen and fibronectin. FEMS Microbiol Lett 206:31–37

    Article  CAS  PubMed  Google Scholar 

  • MacKenzie DA, Jeffers F, Parker ML, Vibert-Vallet A, Bongaerts RJ, Roos S, Walter J, Juge N (2010) Strain-specific diversity of mucus-binding proteins in the adhesion and aggregation properties of Lactobacillus reuteri. Microbiology 156:3368–3378

  • Miyoshi Y, Okada S, Uchimura T, Satoh E (2006) A mucus adhesion promoting protein, MapA, mediates the adhesion of Lactobacillus reuteri to Caco-2 human intestinal epithelial cells. Biosci Biotechnol Biochem 70(7):1622–1628

  • Mukai T, Toba T, Ohori H (1996) Collagen binding of Bifidobacterium adolescentis. Curr Microbiol 34:326–331

    Article  Google Scholar 

  • Munoz MPD, Monedero V (2011) Shotgun phage display of Lactobacillus casei BL23 against collagen and fibronectin. J Microbiol Biotechnol 2:197–203

    Article  Google Scholar 

  • Ouwehand AC, Tolkko S, Salminen S (2001) The effect of digestive enzymes on the adhesion of probiotic bacteria in vitro. J Food Sci 66:856–859

    Article  CAS  Google Scholar 

  • Pérez PF, Minnaard Y, Disalvo EA, De Antoni GL (1998) Surface properties of bifidobacterial strains of human origin. Appl Environ Microbiol 64:21–26

    PubMed Central  PubMed  Google Scholar 

  • Ramiah K, van Reenen CA, Dicks LMT (2009) Expression of the mucus adhesion gene Mub, surface layer protein Slp and adhesion-like factor EF-TU of Lactobacillus acidophilus ATCC 4356 under digestive stress conditions, as monitored with real-time PCR. Probiotics Antimicrob Proteins 1:91–95. doi:10.1007/s12602-009-9009-8

    Article  CAS  Google Scholar 

  • Rodríguez IG, Ruiz L, Gueimonde M, Margolles A, Sanchez B (2012) Factors involved in the colonization and survival of bifidobacteria in the gastrointestinal tract. Microbiol Lett. doi:10.1111/1574-6968.12056

    Google Scholar 

  • Roos S, Jonsson H (2002) A high-molecular-mass cell-surface protein from Lactobacillus reuteri 1063 adheres to mucus components. Microbiology 148:433–442

    CAS  PubMed  Google Scholar 

  • Roos S, Aleljung P, Robert N, Lee B, Wadstrom LM, Jonsson H (1996) A collagen binding protein from Lactobacillus reuteri is part of an ABC transporter system. FEMS Microbiol Lett 144:33–38

    Article  CAS  PubMed  Google Scholar 

  • Schillinger U, Guigas C, Holzapfel WH (2005) In vitro adherence and other properties of lactobacilli used in probiotic yoghurt-like products. Int Dairy J 15:1289–1297

    Article  CAS  Google Scholar 

  • Sillanpaa J, Martínez B, Antikainen J, Toba T, Kalkkinen N, Tankka S, Lounatmaa K (2000) Characterization of the collagen-binding S-layer protein CbsA of Lactobacillus crispatus. J Bacteriol 182:6440–6450

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Styriak I, Nemcova R, Chang YH, Ljungh A (2003) Binding of extracellular matrix molecules by probiotic bacteria. Lett Appl Microbiol 37:329–333

    Article  CAS  PubMed  Google Scholar 

  • Sun Z, Huang L, Kong J, Hu S, Zhang X, Kong W (2012) In vitro evaluation of Lactobacillus crispatus K313 and K243: high-adhesion activity and anti-inflammatory effect on Salmonella braenderup infected intestinal epithelial cell. Vet Microbiol 14:212–220

  • Szilagyi A, Shrier I, Heilpern D, Je J, Park S, Chong G, Lalonde C, Cote LF, Lee B (2010) Differential impact of lactose/lactase phenotype on colonic microflora. Can J Gastroenterol 24:373–379

    PubMed Central  PubMed  Google Scholar 

  • Tallon R, Arias S, Bressollier P, Urdaci MC (2006) Strain and matrix dependent adhesion of Lactobacillus plantarum is mediated by proteinaceous bacterial compounds. J Appl Microbiol 102:442–451

    Google Scholar 

  • Tuomola EM, Ouwehand AC, Salminen S (1999a) Human ileostomy glycoproteins as a model for small intestinal mucus to investigate adhesion of probiotics. Lett Appl Microbiol 28:159–163

    Article  CAS  PubMed  Google Scholar 

  • Tuomola EM, Ouwehand AC, Salminen SJ (1999b) The effect of probiotic bacteria on the adhesion of pathogens to human intestinal mucus. FEMS Immunm Med Microbiol 26:137–142

    Article  CAS  Google Scholar 

  • Tuomola EM, Ouwehand AC, Salminen SJ (2000) Chemical, physical and enzymatic pre-treatments of probiotic lactobacilli alter their adhesion to human intestinal mucus glycoproteins. Int J Food Microbiol 60:75–81

  • Tuomola E, Crittenden R, Playne M, Isolauri E, Salminen S (2001) Quality assurance criteria for probiotics bacteria. Amer J Clin Nutr 73:393–398

    Google Scholar 

  • Turroni F, Serafini F, Foroni E, Duranti S, Motherway MOC, Taverniti V, Mangifesta M, Milani C, Viappiani A, Roversi T, Sánchez B, Santoni A, Gioiosa L, Ferrarini A, Delledonne M, Margolles A, Piazza L, Palanza P, Bolchi A, Guglielmetti S, van Sinderen D, Ventura M (2013) Role of sortase-dependent pili of Bifidobacterium bifidum PRL2010 in modulating bacterium-host interactions. Proc Natl Acad Sci USA 110(27):11151–11156. doi:10.1073/pnas.1303897110

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Velez MP, De Keersmaecker SC, Vanderleyden J (2007) Adherence factors of Lactobacillus in the human gastrointestinal tract. FEMS Microbiol Lett 276:140–148

    Article  CAS  PubMed  Google Scholar 

  • Ventura M, Jankovic I, Walker DC, Pridmore RD, Zink R (2002) Identification and characterization of novel surface proteins in Lactobacillus johnsonii and Lactobacillus gasseri. Appl Environ Microbiol 68:6172–6181

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Vesterlund S, Paltta J, Karp M, Ouwehand AC (2005) Adhesion of bacteria to resected human colonic tissue quantitative analysis of bacterial adhesion and viability. Res Microbiol 56:238–244

    Article  Google Scholar 

  • Vidgren 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 174:7419–7427

    PubMed Central  CAS  PubMed  Google Scholar 

  • Yadav AK, Tyagi A, Kaushik JK, Saklani AC, Grover S, Batish VK (2013) Role of surface layer collagen binding protein from indigenous Lactobacillus plantarum 91 in adhesion and its anti-adhesion potential against gut pathogen. Microbiol Res 168:639–645

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors greatly appreciate the financial support received from Department of Biotechnology, Govt. of India, India.

Conflict of interest

None of the authors have a conflict of interest for the publication of the manuscript.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, National Institute of Nutrition, Hyderabad, India

    Ashok Kumar Yadav

  2. Molecular Biology Unit, Dairy Microbiology Division, National Dairy Research Institute, Karnal, 132001, Haryana, India

    Ashish Tyagi, Sunita Grover & Virender Kumar Batish

  3. Department of Biotechnology, Seth Jai Parkash Mukand Lal Institute of Engineering and Technology, Radaur, Yamuna Nagar, 135133, Haryana, India

    Ashwani Kumar

  4. School of Life Sciences, Appejay Stya University, Gurgaon, Haryana, India

    Asha Chandola Saklani

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  1. Ashok Kumar Yadav
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  2. Ashish Tyagi
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  5. Sunita Grover
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  6. Virender Kumar Batish
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Correspondence to Ashok Kumar Yadav or Virender Kumar Batish.

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Communicated by Erko Stackebrandt.

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Yadav, A.K., Tyagi, A., Kumar, A. et al. Adhesion of indigenous Lactobacillus plantarum to gut extracellular matrix and its physicochemical characterization. Arch Microbiol 197, 155–164 (2015). https://doi.org/10.1007/s00203-014-1034-7

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  • DOI: https://doi.org/10.1007/s00203-014-1034-7

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