DOI QR코드

DOI QR Code

LC-MS/MS Analysis of Surface Layer Proteins as a Useful Method for the Identification of Lactobacilli from the Lactobacillus acidophilus Group

  • Podlesny, Marcin (Department of Biotechnology, Human Nutrition and Food Commodities, University of Life Sciences in Lublin) ;
  • Jarocki, Piotr (Department of Biotechnology, Human Nutrition and Food Commodities, University of Life Sciences in Lublin) ;
  • Komon, Elwira (Department of Biotechnology, Human Nutrition and Food Commodities, University of Life Sciences in Lublin) ;
  • Glibowska, Agnieszka (Department of Biotechnology, Human Nutrition and Food Commodities, University of Life Sciences in Lublin) ;
  • Targonski, Zdzislaw (Department of Biotechnology, Human Nutrition and Food Commodities, University of Life Sciences in Lublin)
  • Received : 2010.09.27
  • Accepted : 2010.12.27
  • Published : 2011.04.28

Abstract

For precise identification of a Lactobacillus K1 isolate, LC-MS/MS analysis of the putative surface layer protein was performed. The results obtained from LTQ-FT-ICR mass spectrometry confirmed that the analyzed protein spot is the surface layer protein originating from Lb. helveticus species. Moreover, the identified protein has the highest similarity with the surface layer protein from Lb. helveticus R0052. To evaluate the proteomic study, multilocus sequence analysis of selected housekeeping gene sequences was performed. Combination of 16S rRNA sequencing with partial sequences for the genes encoding the RNA polymerase alpha subunit (rpoA), phenylalanyl-tRNA synthase alpha subunit (pheS), translational elongation factor Tu (tuf), and Hsp60 chaperonins (groEL) also allowed to classify the analyzed isolate as Lb. helveticus. Further classification at the strain level was achieved by sequencing of the slp gene. This gene showed 99.8% identity with the corresponding slp gene of Lb. helveticus R0052, which is in good agreement with data obtained by nano-HPLC coupled to an LTQ-FT-ICR mass spectrometer. Finally, LC-MS/MS analysis of surface layer proteins extracted from three other Lactobacillus strains proved that the proposed method is the appropriate molecular tool for the identification of S-layer-possessing lactobacilli at the species and even strain levels.

Keywords

References

  1. Anderson, J. W. and S. E. Gilliland. 1999. Effect of fermented milk (yogurt) containing Lactobacillus acidophilus L1 on serum cholesterol in hypercholesterolemic humans. J. Am. Coll. Nutr. 18: 43-50.
  2. Antikainen, J., L. Anton, J. Sillanpaa, and T. K. Korhonen. 2002. Domains in the S-layer protein CbsA of Lactobacillus cispatus involved in adherence to collagens, laminin and lipoteichoic acids and in self-assembly. Mol. Microbiol. 46: 381-394. https://doi.org/10.1046/j.1365-2958.2002.03180.x
  3. Avall-Jaaskelainen, S., U. Hynonen, N. Ilk, D. Pum, U. B. Sleytr, and A. Palva. 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. I8: 165.
  4. Avall-Jaaskelainen, S. and A. Palva. 2005. Lactobacillus surface layers and their applications. FEMS Microbiol. Rev. 29: 511-529.
  5. Axelsson, L. 2004. Lactic acid bacteria: Classification and physiology, pp. 1-66. In S. Salminen, A. von Wright, and A. Ouwehand (eds.). Lactic Acid Bacteria. Microbiological and Functional Aspects, 3rd Ed. Marcel Dekker, New York.
  6. Balcazar, J. L., I. de Blas, I. Ruiz-Zarzuela, D. Vendrell, O. Girones, and J. L. Muzquiz. 2007. Sequencing of variable regions of the 16S rRNA gene for identification of lactic acid bacteria isolated from the intestinal microbiota of healthy salmonids. Comp. Immunol. Microbiol. Infect. Dis. 30: 111-118. https://doi.org/10.1016/j.cimid.2006.12.001
  7. Baumeister, W. and G. Lembcke. 1992. Structural features of archaebacterial cell envelopes. J. Bioenerg. Biomembr. 24: 567- 575. https://doi.org/10.1007/BF00762349
  8. Berger, B., R. D. Pridmore, C. Barretto, F. Delmas-Julien, K. Schreiber, F. Arigoni, and H. Brussow. 2007. Similarity and differences in the Lactobacillus acidophilus group identified by polyphasic analysis and comparative genomics. J. Bacteriol. 189: 1311-1321. https://doi.org/10.1128/JB.01393-06
  9. Boot, H. J., C. P. Kolen, B. Pot, K. Kersters, and P. H. Pouwels. 1996. The presence of two S-layer-protein-encoding genes is conserved among species related to Lactobacillus acidophilus. Microbiology 142: 2375-2384. https://doi.org/10.1099/00221287-142-9-2375
  10. Callegari, M. L., B. Riboli, J. W. Sanders, P. S. Cocconcelli, J. Kok, G. Venema, and L. Morelli. 1998. The S-layer gene of Lactobacillus helveticus CNRZ 892: Cloning, sequence and heterologous expression. Microbiology 144: 719-726. https://doi.org/10.1099/00221287-144-3-719
  11. Chen, X., Y. Chen, X. Li, N. Chen, and W. Fang. 2009. Characterization of surface layer proteins in Lactobacillus crispatus isolate ZJ001. J. Microbiol. Biotechnol. 19: 1176-1183.
  12. Chen, X., J. Xu, J. Shuai, J. Chen, Z. Zhang, and W. Fang. 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. https://doi.org/10.1016/j.ijfoodmicro.2006.11.007
  13. de Moreno de LeBlanc, A., C. Matar, and G. Perdigon. 2007. The application of probiotics in cancer. Br. J. Nutr. 98(Suppl. 1): 105-110.
  14. Egelseer, E. M., I. Schocher, U. B. Sleytr, and M. Sara. 1996. Evidence that an N-terminal S-layer protein fragment triggers the release of a cell-associated high molecular-weight amylase from Bacillus stearothermophilus ATCC 12980. J. Bacteriol. 178: 5602-5609.
  15. Gatti, M., L. Rossetti, M. E. Fornasari, C. Lazzi, G. Giraffa, and E. Neviani. 2005. Heterogeneity of putative surface layer proteins in Lactobacillus helveticus. Appl. Environ. Microbiol. 71: 7582-7588. https://doi.org/10.1128/AEM.71.11.7582-7588.2005
  16. Guarner, F. and G. J. Schaafsma. 1998. Probiotics. Int. J. Food Microbiol. 39: 237-238 https://doi.org/10.1016/S0168-1605(97)00136-0
  17. Hagen, K. E., L. L. Guan, G. W. Tannock, D. R. Korver, and G. E. Allison. 2005. Detection, characterization, and in vitro expression of genes encoding S-proteins in Lactobacillus gallinarum strains isolated from chicken crops. Appl. Environ. Microbiol. 71: 6633- 6643. https://doi.org/10.1128/AEM.71.11.6633-6643.2005
  18. Jahn-Schmid, B., M. Graninger, M. Glozik, S. Kupcu, C. Ebner, F. M. Unger, U. B. Sleytr, and P. Messner. 1996. Immunoreactivity of allergen (Bet v 1) conjugated to crystalline bacterial cell surface layers (S-layers). Immunotechnology 2: 103-113. https://doi.org/10.1016/1380-2933(96)00041-3
  19. Jakava-Viljanen, M., S. Avall-Jaaskelainen, P. Messner, U. B. Sleytr, and A. Palva. 2002. Isolation of three new surface layer protein genes (slp) from Lactobacillus brevis ATCC 14869 and characterization of the change in their expression under aerated and anaerobic conditions. J. Bacteriol. 184: 6786-6795. https://doi.org/10.1128/JB.184.24.6786-6795.2002
  20. Jakava-Viljanen, M. and A. Palva. 2007. Isolation of surface (S) layer protein carrying Lactobacillus species from porcine intestine and faeces and characterization of their adhesion properties to different host tissues. Vet. Microbiol. 124: 264-273. https://doi.org/10.1016/j.vetmic.2007.04.029
  21. Jarocki, P., M. Podlesny, A. Wasko, A. Siuda, and Z. Targonski. 2010. Differentiation of 3 Lactobacillus rhamnosus strains: E/N, Oxy and Pen by SDS-PAGE and two-dimensional electrophoresis of surface-associated proteins. J. Microbiol. Biotechnol. 20: 558-562.
  22. Kim, T-W., H.-S. Song, and H.-Y. Kim. 2005. Distribution of dominant Bifidobacteria in intestinal microflora of Korean adults and seniors, identified by SDS-PAGE of whole cell proteins and 16S rDNA sequence. J. Microbiol. Biotechnol. 15: 388-394.
  23. Kwon, O. S. 2000. Characterization of isolated Lactobacillus spp. and classification by RAPD-PCR analysis. J. Microbiol. 38: 137-144.
  24. Lebeer, S., J. Vanderleyden, and S. C. J. De Keersmaecker. 2008. Genes and molecules of Lactobacilli supporting probiotic action. Microbiol. Mol. Biol. Rev. 72: 728-764. https://doi.org/10.1128/MMBR.00017-08
  25. Markiewicz, L., E. Biedrzycka, and M. Bielecka. 2006. Differentiation of dairy Lactobacillus strains using PFGE method. Food Sci. Technol. Quality. 47(Suppl.): 216-222.
  26. Marteau, P., B. Flourie, P. Pochart, C. Chastang, J. F. Desjeux, and J. C. Rambaud. 1990. Effect of the microbial lactase (E.C. 3.2.1.23) activity in yoghurt on the intestinal absorption of lactose: An in vivo study in lactase-deficient humans. Br. J. Nutr. 64: 71-79. https://doi.org/10.1079/BJN19900010
  27. Matsuzaki, T., R. Yamazaki, S. Hashimoto, and T. Yokokura. 1998. The effect of oral feeding of Lactobacillus casei strain Shirota on immunoglobulin E production in mice. J. Dairy Sci. 81: 48-53. https://doi.org/10.3168/jds.S0022-0302(98)75549-3
  28. Masuda, K. 1992. Heterogeneity of S-layer proteins of Lactobacillus acidophilus strains. Microbiol. Immunol. 36: 297-301.
  29. Miettinen, M., S. Matikainen, J. Vuopio-Varkila, J. Pirhonen, K. Varkila, M. Kurimoto, and I. Julkunen. 1998. Lactobacilli and streptococci induce interleukin-12 (IL-12), IL-18 and gamma interferon production in human peripheral blood mononuclear cells. Infect. Immun. 66: 6058-6062.
  30. Nasser, S. M., K. E. Hagen, M. Vancanneyt, I. Cleenwerck, J. Swings, and T. A. Tompkins. 2006. Lactobacillus suntoryeus Cachat and Priest 2005 is a later synonym of Lactobacillus helveticus (Orla-Jensen 1919) Bergey et al. 1925 (Approved Lists 1980). Int. J. Syst. Evol. Microbiol. 56: 355-360. https://doi.org/10.1099/ijs.0.64001-0
  31. Ouwehand, A. C., E. M. Tuomola, S. Tolkko, and S. Salminen. 2001. Assessment of adhesion properties of novel probiotic strains to human intestinal mucus. Int. J. Food Microbiol. 64: 119-126. https://doi.org/10.1016/S0168-1605(00)00440-2
  32. Parvez, S., K. A. Malik, A. Kang, and H.-Y. Kim. 2006. Probiotics and their fermented food products are beneficial for health. J. Appl. Microbiol. 100: 1171-1185. https://doi.org/10.1111/j.1365-2672.2006.02963.x
  33. Pleschberger, M., A. Neubauer, E. M. Egelseer, S. Weigert, B. Lindner, U. B. Sleytr, S. Muyldermans, and M. Sara. 2003. Generation of a functional monomolecular protein lattice consisting of an S-layer fusion protein comprising the variable domain of a camel heavy chain antibody. Bioconjug. Chem. 14: 440-448. https://doi.org/10.1021/bc025603+
  34. Roberfroid, M. B. 2000. Prebiotics and probiotics: Are they functional foods? Am. J. Clin. Nutr. 71: 1682-1687.
  35. Sampathkumar, P. and M. L. Gilchrist Jr. 2004. Synthesis and characterization of bioconjugates of S-layer proteins. Bioconjug. Chem. 15: 685-693. https://doi.org/10.1021/bc034204r
  36. Sanchez, B., P. Bressollier, and M. C. Urdaci. 2008. Exported proteins in probiotic bacteria: Adhesion to intestinal surfaces, host immunomodulation and molecular cross-talking with the host. FEMS Immunol. Med. Microbiol. 54: 1-17. https://doi.org/10.1111/j.1574-695X.2008.00454.x
  37. Sara, M., D. Pum, and U. B. Sleytr. 1992. Permeability and charge-dependent adsorption properties of the S-layer lattice from Bacillus coagulans E38-66. J. Bacteriol. 174: 3487-3493.
  38. Sara, M. and U. B. Sleytr. 1987. Molecular sieving through Slayers of Bacillus stearothermophilus strains. J. Bacteriol. 169: 4092-4098.
  39. Schneitz, C., L. Nuotio, and K. Lounatma. 1993. Adhesion of Lactobacillus acidophilus to avian intestinal epithelial cells mediated by the crystalline bacterial cell surface layer (S-layer). J. Appl. Bacteriol. 74: 290-294. https://doi.org/10.1111/j.1365-2672.1993.tb03028.x
  40. Sleytr, U. B., C. Huber, N. Ilk, D. Pum, B. Schuster, and E. M. Egelseer. 2007. S-Layers as a tool kit for nanobiotechnological applications. FEMS Microbiol. Lett. 267: 131-144. https://doi.org/10.1111/j.1574-6968.2006.00573.x
  41. Sleytr, U. B., M. Sara, D. Pum, and B. Schuster. 2001. Characterization and use of crystalline bacterial cell surface layers. Prog. Surf. Sci. 68: 231-278. https://doi.org/10.1016/S0079-6816(01)00008-9
  42. Sleytr, U. B. and P. Messner. 1983. Crystalline surface layers on bacteria. Annu. Rev. Microbiol. 37: 311-339. https://doi.org/10.1146/annurev.mi.37.100183.001523
  43. Slover, C. M. and L. Danziger. 2008. Lactobacillus: A review. Clin. Microbiol. Newslett. 30: 23-27. https://doi.org/10.1016/j.clinmicnews.2008.01.006
  44. Smit, E., F. Oling, R. Demel, B. Martinez, and P. H. Pouwels. 2001. The S-layer protein of Lactobacillus acidophilus ATCC 4356: Identication and characterizations of domains responsible for S-protein assembly and cell wall binding. J. Mol. Biol. 305: 245-257. https://doi.org/10.1006/jmbi.2000.4258
  45. Tynkkynen, S., R. Satokari, M. Saarela, T. Mattila-Sandholm, and M. Saxelin. 1999. Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains. Appl. Environ. Microbiol. 65: 3908-3914
  46. Ventura, M., M. L. Callegari, and L. Morelli. 2006. S-Layer gene as a molecular marker for identification of Lactobacillus helveticus. FEMS Microbiol. Lett. 189: 275-279.
  47. Ventura, M., I. Jankovic, D. C. Walker, R. D. Pridmore, and R. Zink. 2002. Identification and characterization of novel surface proteins in Lactobacillus johnsonii and Lactobacillus gasseri. Appl. Environ. Microbiol. 68: 6172-6181. https://doi.org/10.1128/AEM.68.12.6172-6181.2002
  48. Yamamoto, N., T. Shinoda, and T. Takano. 2000. Molecular cloning and sequence analysis of a gene encoding an extracellular proteinase from Lactobacillus helveticus CP790. Biosci. Biotechnol. Biochem. 64: 1217-1222. https://doi.org/10.1271/bbb.64.1217
  49. Yamano, T., M. Tanida, A. Niijima, K. Maeda, N. Okumura, Y. Fukushima, and K. Nagai. 2006. Effects of the probiotics strain Lactobacillus johnsonii strain La1 on autonomic nerves and blood glucose in rats. Life Sci. 79: 1963-1967. https://doi.org/10.1016/j.lfs.2006.06.038

Cited by

  1. A comprehensive post-market review of studies on a probiotic product containing Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011 vol.2, pp.4, 2011, https://doi.org/10.3920/bm2011.0032
  2. Lactobacillus surface layer proteins: structure, function and applications vol.97, pp.12, 2011, https://doi.org/10.1007/s00253-013-4962-2
  3. Lactobacillus kefiri shows inter-strain variations in the amino acid sequence of the S-layer proteins vol.110, pp.4, 2017, https://doi.org/10.1007/s10482-016-0820-4
  4. Amplicon Sequencing of the slpH Locus Permits Culture-Independent Strain Typing of Lactobacillus helveticus in Dairy Products vol.8, pp.None, 2011, https://doi.org/10.3389/fmicb.2017.01380