Imaging of lactic acid bacteria with AFM--elasticity and adhesion maps and their relationship to biological and structural data.

The adhesion of lactic acid bacteria to the intestinal epithelium is one of the most important factors determining probiotic ability of a bacterial strain. Studying bacterial adhesion requires knowledge of the structure and properties of the bacterial surface, which can be studied by atomic force microscopy under native conditions. The observation of the surface topography of bacteria from the species Lactobacillus crispatus, L. helveticus and L. johnsonii shows major differences between bacteria having a crystalline-like protein layer as part of the cell wall and those without such layers. Force volume images calculated into elasticity and adhesion force maps of different bacterial strains show that L. crispatus and L. helveticus have a surface with a homogeneous stiffness with no adhesion events. This is most likely caused by the S-layer, which completely covers the surface of the bacteria. We infer that the absence of adhesion peaks is caused by the semi-crystalline character of such protein layers, in agreement with the results obtained from electron microscopy. Analysis of a number of L. johnsonii strains shows that these bacteria have surface properties which strongly differ from the L. crispatus and L. helveticus strains. For L. johnsonii DMS20533 and L. johnsonii ATCC33200 high adhesion forces are observed, which can be related to a surface rich in polysaccharides. L. johnsonii ATCC332 has lower adhesion forces compared to the other two and, furthermore, the surface topography shows depressions. We suppose that this strain has a surface pattern consisting of crystalline-like proteins alternating with polysaccharide-rich domains. The wide variety in surface properties of lactobacilli could well have wide-ranging implications for food processing and for health benefits.

[1]  T. Mattila-Sandholm,et al.  New functional foods in the treatment of food allergy. , 1999, Annals of medicine.

[2]  H. Busscher,et al.  The influence of subinhibitory concentrations of ampicillin and vancomycin on physico-chemical surface characteristics of Enterococcus faecalis 1131 , 2002 .

[3]  Y. Dufrêne,et al.  Surface properties of microbial cells probed at the nanometre scale with atomic force microscopy , 2000 .

[4]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[5]  S. Salminen,et al.  Adhesion of some probiotic and dairy Lactobacillus strains to Caco-2 cell cultures. , 1998, International journal of food microbiology.

[6]  W. Norde,et al.  X-ray photoelectron spectroscopy analysis of whole cells and isolated cell walls of gram-positive bacteria: comparison with biochemical analysis , 1997, Journal of bacteriology.

[7]  M. Walch,et al.  Effect of streptolysin O on the microelasticity of human platelets analyzed by atomic force microscopy. , 2000, Ultramicroscopy.

[8]  Darrell H. Reneker,et al.  CHARACTERIZATION OF POLYMER SURFACES WITH ATOMIC FORCE MICROSCOPY , 1997 .

[9]  Miss A.O. Penney (b) , 1974, The New Yale Book of Quotations.

[10]  S. Lortal,et al.  X-ray Photoelectron-spectroscopy and Biochemical-analysis of the Surface of Lactobacillus-helveticus Atcc-12046 , 1995 .

[11]  J. Heijenoort,et al.  S-layer of Lactobacillus helveticus ATCC 12046: isolation, chemical characterization and re-formation after extraction with lithium chloride , 1992 .

[12]  S. Salminen,et al.  Adhesion of four Bifidobacterium strains to human intestinal mucus from subjects in different age groups. , 1999, FEMS microbiology letters.

[13]  Sverre Myhra,et al.  Determination of the spring constants of probes for force microscopy/spectroscopy , 1996 .

[14]  Y. Dufrêne,et al.  X-ray photoelectron spectroscopy analysis of the surface composition of Azospirillum brasilense in relation to growth conditions , 1996 .

[15]  D. Brassart,et al.  Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria. , 1994, Gut.

[16]  T. Beveridge Ultrastructure, chemistry, and function of the bacterial wall. , 1981, International review of cytology.

[17]  Ericka Stricklin-Parker,et al.  Ann , 2005 .

[18]  Andrew G. Glen,et al.  APPL , 2001 .

[19]  S. Salminen,et al.  The ability of probiotic bacteria to bind to human intestinal mucus. , 1998, FEMS microbiology letters.

[20]  H. C. van der Mei,et al.  Correlation between hydrophobicity and resistance to nonoxynol-9 and vancomycin for urogenital isolates of lactobacilli. , 1992, FEMS microbiology letters.

[21]  E. Schiffrin,et al.  Immunomodulation of human blood cells following the ingestion of lactic acid bacteria. , 1995, Journal of dairy science.

[22]  Richard M. Pashley,et al.  Direct measurement of colloidal forces using an atomic force microscope , 1991, Nature.

[23]  M. Curk,et al.  Fourier transform infrared (FTIR) spectroscopy for identifying Lactobacillus species , 1994 .

[24]  G. Dietler,et al.  Force-distance curves by atomic force microscopy , 1999 .

[25]  Drechsler,et al.  A cantilever array-based artificial nose , 2000, Ultramicroscopy.

[26]  A. Engel,et al.  Charting and unzipping the surface layer of Corynebacterium glutamicum with the atomic force microscope , 2002, Molecular microbiology.

[27]  W F Heinz,et al.  Relative microelastic mapping of living cells by atomic force microscopy. , 1998, Biophysical journal.

[28]  Matthias Rief,et al.  Single Molecule Force Spectroscopy on Polysaccharides by Atomic Force Microscopy , 1997, Science.

[29]  H. C. van der Mei,et al.  Probing molecular interactions and mechanical properties of microbial cell surfaces by atomic force microscopy. , 2001, Ultramicroscopy.

[30]  N. Kalkkinen,et al.  A Collagen-Binding S-Layer Protein in Lactobacillus crispatus , 1995, Applied and environmental microbiology.

[31]  P. C. Hiemenz,et al.  Principles of colloid and surface chemistry , 1977 .

[32]  M. Hegner,et al.  Specific antigen/antibody interactions measured by force microscopy. , 1996, Biophysical journal.

[33]  Potassium-selective atomic force microscopy on ion-releasing substrates and living cells. , 2002, Analytical chemistry.

[34]  P. Albertsson,et al.  Partition of Cell Particles and Macromolecules , 1986 .

[35]  FTIR spectroscopy and taxonomic purpose: Contribution to the classification of lactic acid bacteria , 2001 .