1887

Abstract

Structural components of the cell surface have an impact on some of the beneficial attributes of probiotic bacteria. analysis of the NCFM genome sequence revealed the presence of a putative cell surface protein that was predicted to be a myosin cross-reactive antigen (MCRA). As MCRAs are conserved among many probiotic bacteria, we used the -based counterselective gene replacement system, designed recently for use in , to determine the functional role of this gene (LBA649) in NCFM. Phenotypic assays were undertaken with the parent strain (NCK1909) and deletion mutant (NCK2015) to assign a function for this gene. The growth of NCK2015 (ΔLBA649) was reduced in the presence of lactate, acetate, porcine bile and salt. Adhesion of NCK2015 to Caco-2 cells was substantially reduced for both stationary-phase (∼45 % reduction) and exponential-phase cells (∼50 % reduction). Analysis of NCK2015 by scanning electron microscopy revealed a longer cell morphology after growth in MRS broth compared to NCK1909. These results indicate a role for LBA649 in stress tolerance, cell wall division and adherence to Caco-2 cells.

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2010-11-01
2019-10-15
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References

  1. Altermann, E., Russell, W. M., Azcarate-Peril, M. A., Barrangou, R., Buck, B. L., McAuliffe, O., Souther, N., Dobson, A., Duong, T. & other authors ( 2005; ). Complete genome sequence of the probiotic lactic acid bacterium Lactobacillus acidophilus NCFM. Proc Natl Acad Sci U S A 102, 3906–3912.[CrossRef]
    [Google Scholar]
  2. Altschul, S. F. & Lipman, D. J. ( 1990; ). Protein database searches for multiple alignments. Proc Natl Acad Sci U S A 87, 5509–5513.[CrossRef]
    [Google Scholar]
  3. Azcarate-Peril, M. A., Altermann, E., Hoover-Fitzula, R. L., Cano, R. J. & Klaenhammer, T. R. ( 2004; ). Identification and inactivation of genetic loci involved with Lactobacillus acidophilus acid tolerance. Appl Environ Microbiol 70, 5315–5322.[CrossRef]
    [Google Scholar]
  4. Azcarate-Peril, M. A., McAuliffe, O., Altermann, E., Lick, S., Russell, W. M. & Klaenhammer, T. R. ( 2005; ). Microarray analysis of a two-component regulatory system involved in acid resistance and proteolytic activity in Lactobacillus acidophilus. Appl Environ Microbiol 71, 5794–5804.[CrossRef]
    [Google Scholar]
  5. Azcarate-Peril, M. A., Bruno-Barcena, J. M., Hassan, H. M. & Klaenhammer, T. R. ( 2006; ). Transcriptional and functional analysis of oxalyl-coenzyme A (CoA) decarboxylase and formyl-CoA transferase genes from Lactobacillus acidophilus. Appl Environ Microbiol 72, 1891–1899.[CrossRef]
    [Google Scholar]
  6. Barrangou, R., Altermann, E., Hutkins, R., Cano, R. & Klaenhammer, T. R. ( 2003; ). Functional and comparative genomic analyses of an operon involved in fructooligosaccharide utilization by Lactobacillus acidophilus. Proc Natl Acad Sci U S A 100, 8957–8962.[CrossRef]
    [Google Scholar]
  7. Barrangou, R., Azcarate-Peril, M. A., Duong, T., Conners, S. B., Kelly, R. M. & Klaenhammer, T. R. ( 2006; ). Global analysis of carbohydrate utilization by Lactobacillus acidophilus using cDNA microarrays. Proc Natl Acad Sci U S A 103, 3816–3821.[CrossRef]
    [Google Scholar]
  8. Båth, K., Roos, S., Wall, T. & Jonsson, H. ( 2005; ). The cell surface of Lactobacillus reuteri ATCC 55730 highlighted by identification of 126 extracellular proteins from the genome sequence. FEMS Microbiol Lett 253, 75–82.[CrossRef]
    [Google Scholar]
  9. Begley, M., Gahan, C. G. & Hill, C. ( 2005; ). The interaction between bacteria and bile. FEMS Microbiol Rev 29, 625–651.[CrossRef]
    [Google Scholar]
  10. Bendtsen, J. D., Nielsen, H., von Heijne, G. & Brunak, S. ( 2004; ). Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340, 783–795.[CrossRef]
    [Google Scholar]
  11. Bevers, L. E., Pinkse, M. W., Verhaert, P. D. & Hagen, W. R. ( 2009; ). Oleate hydratase catalyzes the hydration of a nonactivated carbon–carbon bond. J Bacteriol 191, 5010–5012.[CrossRef]
    [Google Scholar]
  12. Boneca, I. G., Dussurget, O., Cabanes, D., Nahori, M. A., Sousa, S., Lecuit, M., Psylinakis, E., Bouriotis, V., Hugot, J. P. & other authors ( 2007; ). A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system. Proc Natl Acad Sci U S A 104, 997–1002.[CrossRef]
    [Google Scholar]
  13. Buck, B. L., Altermann, E., Svingerud, T. & Klaenhammer, T. R. ( 2005; ). Functional analysis of putative adhesion factors in Lactobacillus acidophilus NCFM. Appl Environ Microbiol 71, 8344–8351.[CrossRef]
    [Google Scholar]
  14. Buck, B. L., Azcarate-Peril, M. A. & Klaenhammer, T. R. ( 2009; ). Role of autoinducer-2 on the adhesion ability of Lactobacillus acidophilus. J Appl Microbiol 107, 269–279.[CrossRef]
    [Google Scholar]
  15. Claros, M. G. & von Heijne, G. ( 1994; ). TopPred II: an improved software for membrane protein structure predictions. Comput Appl Biosci 10, 685–686.
    [Google Scholar]
  16. Coakley, M., Ross, R. P., Nordgren, M., Fitzgerald, G., Devery, R. & Stanton, C. ( 2003; ). Conjugated linoleic acid biosynthesis by human-derived Bifidobacterium species. J Appl Microbiol 94, 138–145.[CrossRef]
    [Google Scholar]
  17. Cserzö, M., Eisenhaber, F., Eisenhaber, B. & Simon, I. ( 2002; ). On filtering false positive transmembrane protein predictions. Protein Eng 15, 745–752.[CrossRef]
    [Google Scholar]
  18. Dobson, A. E., Sanozky-Dawes, R. B. & Klaenhammer, T. R. ( 2007; ). Identification of an operon and inducing peptide involved in the production of lactacin B by Lactobacillus acidophilus. J Appl Microbiol 103, 1766–1778.[CrossRef]
    [Google Scholar]
  19. Fischer, H., Yamamoto, M., Akira, S., Beutler, B. & Svanborg, C. ( 2006; ). Mechanism of pathogen-specific TLR4 activation in the mucosa: fimbriae, recognition receptors and adaptor protein selection. Eur J Immunol 36, 267–277.[CrossRef]
    [Google Scholar]
  20. Goh, Y. J., Azcarate-Peril, M. A., O'Flaherty, S., Durmaz, E., Valence, F., Jardin, J., Lortal, S. & Klaenhammer, T. R. ( 2009; ). Development and application of a upp-based counterselective gene replacement system for the study of the S-layer protein SlpX of Lactobacillus acidophilus NCFM. Appl Environ Microbiol 75, 3093–3105.[CrossRef]
    [Google Scholar]
  21. Gopal, P. K., Prasad, J., Smart, J. & Gill, H. S. ( 2001; ). In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli. Int J Food Microbiol 67, 207–216.[CrossRef]
    [Google Scholar]
  22. Grangette, C., Nutten, S., Palumbo, E., Morath, S., Hermann, C., Dewulf, J., Pot, B., Hartung, T., Hols, P. & Mercenier, A. ( 2005; ). Enhanced antiinflammatory capacity of a Lactobacillus plantarum mutant synthesizing modified teichoic acids. Proc Natl Acad Sci U S A 102, 10321–10326.[CrossRef]
    [Google Scholar]
  23. Hanahan, D. ( 1985; ). Techniques for transformation of E. coli. In DNA Cloning: a Practical Approach, pp. 109–135. Edited by Glover, D. M.. Oxford, UK. : IRL Press Ltd.
    [Google Scholar]
  24. Jenkins, J. K. & Courtney, P. D. ( 2003; ). Lactobacillus growth and membrane composition in the presence of linoleic or conjugated linoleic acid. Can J Microbiol 49, 51–57.[CrossRef]
    [Google Scholar]
  25. Kil, K. S., Cunningham, M. W. & Barnett, L. A. ( 1994; ). Cloning and sequence analysis of a gene encoding a 67-kilodalton myosin-cross-reactive antigen of Streptococcus pyogenes reveals its similarity with class II major histocompatibility antigens. Infect Immun 62, 2440–2449.
    [Google Scholar]
  26. Kingsford, C. L., Ayanbule, K. & Salzberg, S. L. ( 2007; ). Rapid, accurate, computational discovery of Rho-independent transcription terminators illuminates their relationship to DNA uptake. Genome Biol 8, R22.[CrossRef]
    [Google Scholar]
  27. Konstantinov, S. R., Smidt, H., de Vos, W. M., Bruijns, S. C., Singh, S. K., Valence, F., Molle, D., Lortal, S., Altermann, E. & other authors ( 2008; ). S layer protein A of Lactobacillus acidophilus NCFM regulates immature dendritic cell and T cell functions. Proc Natl Acad Sci U S A 105, 19474–19479.[CrossRef]
    [Google Scholar]
  28. Krogh, A., Larsson, B., von Heijne, G. & Sonnhammer, E. L. ( 2001; ). Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. J Mol Biol 305, 567–580.[CrossRef]
    [Google Scholar]
  29. Law, J., Buist, G., Haandrikman, A., Kok, J., Venema, G. & Leenhouts, K. ( 1995; ). A system to generate chromosomal mutations in Lactococcus lactis which allows fast analysis of targeted genes. J Bacteriol 177, 7011–7018.
    [Google Scholar]
  30. Lebeer, S., Vanderleyden, J. & De Keersmaecker, S. C. ( 2010; ). Host interactions of probiotic bacterial surface molecules: comparison with commensals and pathogens. Nat Rev Microbiol 8, 171–184.[CrossRef]
    [Google Scholar]
  31. Leyer, G. J., Li, S., Mubasher, M. E., Reifer, C. & Ouwehand, A. C. ( 2009; ). Probiotic effects on cold and influenza-like symptom incidence and duration in children. Pediatrics 124, e172–e179.[CrossRef]
    [Google Scholar]
  32. Liévin-Le Moal, V., Sarrazin-Davila, L. E. & Servin, A. L. ( 2007; ). An experimental study and a randomized, double-blind, placebo-controlled clinical trial to evaluate the antisecretory activity of Lactobacillus acidophilus strain LB against nonrotavirus diarrhea. Pediatrics 120, e795–e803.[CrossRef]
    [Google Scholar]
  33. Lin, C. K., Tsai, H. C., Lin, P. P., Tsen, H. Y. & Tsai, C. C. ( 2008; ). Lactobacillus acidophilus LAP5 able to inhibit the Salmonella choleraesuis invasion to the human Caco-2 epithelial cell. Anaerobe 14, 251–255.[CrossRef]
    [Google Scholar]
  34. Mohamadzadeh, M. & Klaenhammer, T. R. ( 2008; ). Specific Lactobacillus species differentially activate Toll-like receptors and downstream signals in dendritic cells. Expert Rev Vaccines 7, 1155–1164.[CrossRef]
    [Google Scholar]
  35. O'Flaherty, S. & Klaenhammer, T. R. ( 2010; ). The role and potential of probiotic bacteria in the gut, and the communication between gut microflora and gut/host. Int Dairy J 20, 262–268.[CrossRef]
    [Google Scholar]
  36. Pfeiler, E. A. & Klaenhammer, T. R. ( 2009; ). Role of transporter proteins in bile tolerance of Lactobacillus acidophilus. Appl Environ Microbiol 75, 6013–6016.[CrossRef]
    [Google Scholar]
  37. Pfeiler, E. A., Azcarate-Peril, M. A. & Klaenhammer, T. R. ( 2007; ). Characterization of a novel bile-inducible operon encoding a two-component regulatory system in Lactobacillus acidophilus. J Bacteriol 189, 4624–4634.[CrossRef]
    [Google Scholar]
  38. Quevillon, E., Silventoinen, V., Pillai, S., Harte, N., Mulder, N., Apweiler, R. & Lopez, R. ( 2005; ). InterProScan: protein domains identifier. Nucleic Acids Res 33, W116–W120.
    [Google Scholar]
  39. Roos, S., Engstrand, L. & Jonsson, H. ( 2005; ). Lactobacillus gastricus sp. nov., Lactobacillus antri sp. nov., Lactobacillus kalixensis sp. nov. and Lactobacillus ultunensis sp. nov., isolated from human stomach mucosa. Int J Syst Evol Microbiol 55, 77–82.[CrossRef]
    [Google Scholar]
  40. Rousseaux, C., Thuru, X., Gelot, A., Barnich, N., Neut, C., Dubuquoy, L., Dubuquoy, C., Merour, E., Geboes, K. & other authors ( 2007; ). Lactobacillus acidophilus modulates intestinal pain and induces opioid and cannabinoid receptors. Nat Med 13, 35–37.[CrossRef]
    [Google Scholar]
  41. Russell, W. M. & Klaenhammer, T. R. ( 2001; ). Efficient system for directed integration into the Lactobacillus acidophilus and Lactobacillus gasseri chromosomes via homologous recombination. Appl Environ Microbiol 67, 4361–4364.[CrossRef]
    [Google Scholar]
  42. Sambrook, J. & Russel, D. W. ( 2001; ). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY. : Cold Spring Harbor Laboratory.
    [Google Scholar]
  43. Tusnády, G. E. & Simon, I. ( 2001; ). The hmmtop transmembrane topology prediction server. Bioinformatics 17, 849–850.[CrossRef]
    [Google Scholar]
  44. van Pijkeren, J. P., Canchaya, C., Ryan, K. A., Li, Y., Claesson, M. J., Sheil, B., Steidler, L., O'Mahony, L., Fitzgerald, G. F. & other authors ( 2006; ). Comparative and functional analysis of sortase-dependent proteins in the predicted secretome of Lactobacillus salivarius UCC118. Appl Environ Microbiol 72, 4143–4153.[CrossRef]
    [Google Scholar]
  45. Volkov, A., Liavonchanka, A., Kamneva, O., Fiedler, T., Goebel, C., Kreikemeyer, B. & Feussner, I. ( 2010; ). Myosin cross-reactive antigen of Streptococcus pyogenes M49 encodes a fatty acid double bond hydratase that plays a role in oleic acid detoxification and bacterial virulence. J Biol Chem 285, 10353–10361.[CrossRef]
    [Google Scholar]
  46. Walker, D. C., Aoyama, K. & Klaenhammer, T. R. ( 1996; ). Electrotransformation of Lactobacillus acidophilus group A1. FEMS Microbiol Lett 138, 233–237.[CrossRef]
    [Google Scholar]
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