Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Feb;176(3):848–860. doi: 10.1128/jb.176.3.848-860.1994

Isolation of two physiologically induced variant strains of Bacillus stearothermophilus NRS 2004/3a and characterization of their S-layer lattices.

M Sára 1, D Pum 1, S Küpcü 1, P Messner 1, U B Sleytr 1
PMCID: PMC205123  PMID: 8300538

Abstract

During growth of Bacillus stearothermophilus NRS 2004/3a in continuous culture on complex medium, the chemical properties of the S-layer glycoprotein and the characteristic oblique lattice were maintained only if glucose was used as the sole carbon source. With increased aeration, amino acids were also metabolized, accompanied by liberation of ammonium and by changes in the S-layer protein. Depending on the stage of fermentation at which oxygen limitation was relieved, two different variants, one with a more delicate oblique S-layer lattice (variant 3a/V1) and one with a square S-layer lattice (variant 3a/V2), were isolated. During the switch from the wild-type strain to a variant or from variant 3a/V2 to variant 3a/V1, monolayers of two types of S-layer lattices could be demonstrated on the surfaces of single cells. S-layer proteins from variants had different molecular sizes and a significantly lower carbohydrate content than S-layer proteins from the wild-type strain did. Although the S-layer lattices from the wild-type and variant strains showed quite different protein mass distributions in two- and three-dimensional reconstructions, neither the amino acid composition nor the pore size, as determined by permeability studies, was significantly changed. Peptide mapping and N-terminal sequencing results strongly indicated that the three S-layer proteins are encoded by different genes and are not derived from a universal precursor form.

Full text

PDF
857

Images in this article

851Image
on p.851
852Image
on p.852
853Image
on p.853
854Image
on p.854
856Image
on p.856
857Image
on p.857
857Image
on p.857
858Image
on p.858

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amos L. A., Henderson R., Unwin P. N. Three-dimensional structure determination by electron microscopy of two-dimensional crystals. Prog Biophys Mol Biol. 1982;39(3):183–231. doi: 10.1016/0079-6107(83)90017-2. [DOI] [PubMed] [Google Scholar]
  2. Beveridge T. J., Graham L. L. Surface layers of bacteria. Microbiol Rev. 1991 Dec;55(4):684–705. doi: 10.1128/mr.55.4.684-705.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breitwieser A., Gruber K., Sleytr U. B. Evidence for an S-layer protein pool in the peptidoglycan of Bacillus stearothermophilus. J Bacteriol. 1992 Dec;174(24):8008–8015. doi: 10.1128/jb.174.24.8008-8015.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Christian R., Schulz G., Unger F. M., Messner P., Küpcü Z., Sleytr U. B. Structure of a rhamnan from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a. Carbohydr Res. 1986 Aug 1;150:265–272. doi: 10.1016/0008-6215(86)80021-0. [DOI] [PubMed] [Google Scholar]
  5. FRANCOIS C., MARSHALL R. D., NEUBERGER A. Carbohydrates in protein. 4. The determination of mannose in hen's-egg albumin by radioisotope dilution. Biochem J. 1962 May;83:335–341. doi: 10.1042/bj0830335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujimoto S., Takade A., Amako K., Blaser M. J. Correlation between molecular size of the surface array protein and morphology and antigenicity of the Campylobacter fetus S layer. Infect Immun. 1991 Jun;59(6):2017–2022. doi: 10.1128/iai.59.6.2017-2022.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hovmöller S., Sjögren A., Wang D. N. The structure of crystalline bacterial surface layers. Prog Biophys Mol Biol. 1988;51(2):131–163. doi: 10.1016/0079-6107(88)90012-0. [DOI] [PubMed] [Google Scholar]
  8. Howard L., Tipper D. J. A polypeptide bacteriophage receptor: modified cell wall protein subunits in bacteriophage-resistant mutants of Bacillus sphaericus strain P-1. J Bacteriol. 1973 Mar;113(3):1491–1504. doi: 10.1128/jb.113.3.1491-1504.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Küpcü Z., März L., Messner P., Sleytr U. B. Evidence for the glycoprotein nature of the crystalline cell wall surface layer of Bacillus stearothermophilus strain NRS2004/3a. FEBS Lett. 1984 Jul 23;173(1):185–190. doi: 10.1016/0014-5793(84)81043-1. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Messner P., Christian R., Kolbe J., Schulz G., Sleytr U. B. Analysis of a novel linkage unit of O-linked carbohydrates from the crystalline surface layer glycoprotein of Clostridium thermohydrosulfuricum S102-70. J Bacteriol. 1992 Apr;174(7):2236–2240. doi: 10.1128/jb.174.7.2236-2240.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Messner P., Pum D., Sleytr U. B. Characterization of the ultrastructure and the self-assembly of the surface layer of Bacillus stearothermophilus strain NRS 2004/3a. J Ultrastruct Mol Struct Res. 1986 Oct-Dec;97(1-3):73–88. doi: 10.1016/s0889-1605(86)80008-8. [DOI] [PubMed] [Google Scholar]
  13. Messner P., Sleytr U. B., Christian R., Schulz G., Unger F. M. Isolation and structure determination of a diacetamidodideoxyuronic acid-containing glycan chain from the S-layer glycoprotein of Bacillus stearothermophilus NRS 2004/3a. Carbohydr Res. 1987 Nov 1;168(2):211–218. doi: 10.1016/0008-6215(87)80027-7. [DOI] [PubMed] [Google Scholar]
  14. Messner P., Sleytr U. B. Crystalline bacterial cell-surface layers. Adv Microb Physiol. 1992;33:213–275. doi: 10.1016/s0065-2911(08)60218-0. [DOI] [PubMed] [Google Scholar]
  15. Pum D., Sára M., Sleytr U. B. Structure, surface charge, and self-assembly of the S-layer lattice from Bacillus coagulans E38-66. J Bacteriol. 1989 Oct;171(10):5296–5303. doi: 10.1128/jb.171.10.5296-5303.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. RONDLE C. J., MORGAN W. T. The determination of glucosamine and galactosamine. Biochem J. 1955 Dec;61(4):586–589. doi: 10.1042/bj0610586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sleytr U. B., Messner P. Crystalline surface layers in procaryotes. J Bacteriol. 1988 Jul;170(7):2891–2897. doi: 10.1128/jb.170.7.2891-2897.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sleytr U. B., Messner P. Crystalline surface layers on bacteria. Annu Rev Microbiol. 1983;37:311–339. doi: 10.1146/annurev.mi.37.100183.001523. [DOI] [PubMed] [Google Scholar]
  19. Sleytr U. B. Self-assembly of the hexagonally and tetragonally arranged subunits of bacterial surface layers and their reattachment to cell walls. J Ultrastruct Res. 1976 Jun;55(3):360–377. doi: 10.1016/s0022-5320(76)80093-7. [DOI] [PubMed] [Google Scholar]
  20. Sleytr U. B., Sára M., Küpcü Z., Messner P. Structural and chemical characterization of S-layers of selected strains of Bacillus stearothermophilus and Desulfotomaculum nigrificans. Arch Microbiol. 1986 Oct;146(1):19–24. doi: 10.1007/BF00690152. [DOI] [PubMed] [Google Scholar]
  21. Sára M., Pum D., Sleytr U. B. Permeability and charge-dependent adsorption properties of the S-layer lattice from Bacillus coagulans E38-66. J Bacteriol. 1992 Jun;174(11):3487–3493. doi: 10.1128/jb.174.11.3487-3493.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sára M., Sleytr U. B. Molecular sieving through S layers of Bacillus stearothermophilus strains. J Bacteriol. 1987 Sep;169(9):4092–4098. doi: 10.1128/jb.169.9.4092-4098.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sørensen S. B., Sørensen T. L., Breddam K. Fragmentation of proteins by S. aureus strain V8 protease. Ammonium bicarbonate strongly inhibits the enzyme but does not improve the selectivity for glutamic acid. FEBS Lett. 1991 Dec 9;294(3):195–197. doi: 10.1016/0014-5793(91)80667-r. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES