1887

Abstract

Isolates of thermophilic bacteria obtained from an Austrian beet sugar factory were screened by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and freeze-fracture electron microscopy for the presence of glycosylated crystalline cell surface layers (S-layers). On the basis of similarities in the protein band patterns on SDS-PAGE gels and the lattice geometry of the S-layers as revealed by electron micrographs, the 31 isolates which we studied were clustered into five groups (groups I to V) and several strains which exhibited no common characteristics (group 0). We found that the organisms belonging to groups I to III had glycosylated S-layer proteins, but the highest carbohydrate contents were observed in group III organisms. Partial sequencing of the 16S ribosomal DNAs of selected representative strains of each group revealed that the group I, II, IV, and V isolates and the few group 0 strains were different from the group III strains. The results of DNA-DNA hybridization experiments, SDS-PAGE, and an analysis of polar lipids demonstrated that group III isolates L419-91, L420-91 (T = type strain), and L438-91 belong to the same species. We chose the group III organism sp. strain L420-91 for further analysis because of the high carbohydrate content of its S-layer protein. The taxonomic position of this isolate was determined by using a polyphasic approach. Phenotypic, chemotaxonomic, and genomic analyses revealed that strains L420-91, L419-91, and L438-91 represent a new species. We observed high levels of similarity between these strains and ATCC 12990, which also had a glycosylated S-layer protein. Our results show that strains L420-91, L419-91, and L438-91 and ATCC 12990 belong to the same species and that this species is a new species, which we name . The type strain of this species is strain L420-91 (= DSM 10154).

Loading

Article metrics loading...

/content/journal/ijsem/10.1099/00207713-46-2-532
1996-04-01
2024-12-05
Loading full text...

Full text loading...

/deliver/fulltext/ijsem/46/2/ijs-46-2-532.html?itemId=/content/journal/ijsem/10.1099/00207713-46-2-532&mimeType=html&fmt=ahah

References

  1. Abe M., Nishimune T., Ito S., Kimoto M., Hayashi R. 1986; A simple method for the detection of two types of thiaminase-producing colonies. FEMS Microbiol. Lett 34:129–133
    [Google Scholar]
  2. Ash C., Farrow J. A. E., Wallbanks S., Collins M. D. 1991; Phylogenetic heterogeneity of the genus Bacillus revealed by comparative analysis of small-subunit-ribosomal RNA sequences. Lett. AppL Microbiol 13:202–206
    [Google Scholar]
  3. Beveridge T. J. 1994; Bacterial S-layers. Curr. Opin. Struct. Biol 4:204–212
    [Google Scholar]
  4. Bligh E. G., Dyer W. J. 1959; A rapid method of total lipid extraction and purification. Biochem. J 83:606–614
    [Google Scholar]
  5. Bock K., Schuster-Kolbe J., Altman E., Allmaier G., Stahl B., Christian R., Sleytr U. B., Messner P. 1994; Primary structure of the O-glycosidically linked glycan chain of the crystalline surface layer glycoprotein of Thermoanaerobacter thermohydrosulfuricus Lil 1-69. Galactosyl tyrosine as a novel linkage unit. J. Biol. Chem 269:7137–7144
    [Google Scholar]
  6. Busse J., Auling G. 1988; Polyamine pattern as a chemotaxonomic marker within the Proteobacteria. Syst. Appl. Microbiol 11:1–8
    [Google Scholar]
  7. Cashion P., Holder-Franklin M. A., McCully J., Franklin M. 1977; A rapid method for the base rate determination of bacterial DNA. Anal. Biochem 81:461–466
    [Google Scholar]
  8. De Ley J., Cattoir H., Reynaerts A. 1970; The quantitative measurement of DNA hybridization from renaturation rates. Eur. J. Biochem 12:133–142
    [Google Scholar]
  9. DeSoete G. 1983; A least squares algorithm for fitting additive trees to proximity data. Psychometrica 48:621–626
    [Google Scholar]
  10. Deutsche Sammlung von Mikroorganismen und Zellkulturen 1993 DSM catalogue of strains. , 5th. Deutsche Sammlung von Mikroorganismen und Zellkulturen; Braunschweig, Germany:
    [Google Scholar]
  11. Dubourg J., Devillers P. 1953; Mikrobeninfektionen in Zuckerfabriken. Z. Zuckerind 3:371–377
    [Google Scholar]
  12. Gordon R. E., Haynes W. C., Pang C. H. 1973; The genus Bacillus. U. S. Dep. Agric. Agric. Handb 427:224–227
    [Google Scholar]
  13. Hamana K., Akiba T., Uchino F., Matsuzaki S. 1989; Distribution of spermine in bacilli and lactic bacteria. Can. J. Microbiol 35:450–455
    [Google Scholar]
  14. Hamana K., Matsuzaki S. 1992; Polyamines as a chemotaxonomic marker in bacterial systematics. Crit. Rev. Microbiol 18:261–283
    [Google Scholar]
  15. Hollaus F., Klaushofer H. 1973; Identification of hyperthermophilic obligate anaerobic bacteria from extraction juices of beet sugar factories. Int. Sugar J 75:237–241
    [Google Scholar]
  16. Hollaus F., Sleytr U. 1972; On the taxonomy and fine structure of some hyperthermophilic saccharolytic Clostridia. Arch. Mikrobiol 86:129–146
    [Google Scholar]
  17. Huss V. A. R., Festl H., Schleifer K. H. 1983; Studies on the spectrophotometric determination of DNA hybridization from renaturation rates. Syst. Appl. Microbiol 4:184–192
    [Google Scholar]
  18. Janke K.-D. 1992; BASIC computer program for evaluation of spectroscopic DNA renaturation data from GILFORD SYSTEM 2600 spectrophotometer on a PC/XT/AT type personal computer. J. Microbiol. Methods 15:61–73
    [Google Scholar]
  19. Jukes T. H., Cantor C. R. 1969 Evolution of protein molecules. 21–132 Munro H. N.ed Mammalian protein metabolism Academic Press, Inc.; New York:
    [Google Scholar]
  20. Kosma P., Neuninger C., Christian R., Schulz G., Messner P. 1995; Glycan structure of the S-layer glycoprotein of Bacillus sp. L420-91. Glycoconjugate J 12:99–107
    [Google Scholar]
  21. Kosma P., Wugeditsch T., Christian R., Messner P. 1995; Glycan structure of a heptose-containing S-layer glycoprotein of Bacillus thermoaerophilus. Glycobiology 5:791–796
    [Google Scholar]
  22. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685
    [Google Scholar]
  23. Lanyi B. 1987; Classical and rapid identification methods for medically important bacteria. Methods Microbiol 19:1–67
    [Google Scholar]
  24. Meier-Stauffer K., Kuen B., Busse H.-J., Hollaus F., Lubitz W., Sleytr U. B., Messner P. 1994 Classification of two thermophilic Bacillus strains based on 16S rDNA and chemotaxonomic analyses. 226 abstr. BC-1/30 Proceedings of the 7th International Congress of Bacteriology and Applied Microbiology Prague; Czech Republic:
    [Google Scholar]
  25. Mesbah M., Premachandran U., Whitman W. B. 1989; Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int. J. Syst, BacterioL 39:159–167
    [Google Scholar]
  26. Messner P. 1996 Chemical composition and biosynthesis of S-layers. 35–76 Sleytr U. B., Messner P., Pum D., Sara M.ed Crystalline bacterial cell surface proteins R. G. Landes Co.; Austin, Tex:
    [Google Scholar]
  27. Messner P., Hollaus F., Sleytr U. B. 1984; Paracrystalline cell wall surface layers of different Bacillus stearothermophilus strains. Int. J. Syst. Bacterio1 34:202–210
    [Google Scholar]
  28. Messner P., Sleytr U. B. 1991; Bacterial surface layer glycoproteins. Glycobiology 1:545–551
    [Google Scholar]
  29. Messner P., Sleytr U. B. 1992; Crystalline bacterial cell-surface layers. Adv. Microb. Physiol 33:213–275
    [Google Scholar]
  30. Minnikin D. E., O’onnell A. G., Goodfellow M., Alderson G., Athalye M., Schaal A., Parlett J. H. 1984; An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids. J. Microbiol. Methods 2:233–241
    [Google Scholar]
  31. Neuninger C. Unpublished data
  32. Podsulo J. F. 1981; Glycoprotein molecular-weight estimation using sodium dodecyl sulfate-pore gradient electrophoresis: comparison of Tris-glycine and Tris-borate-EDTA buffer systems. Anal. Biochem 114:131–139
    [Google Scholar]
  33. Rainey F. A., Dorsch M., Morgan W. H., Stackebrandt E. 1992; 16S rDNA analysis of Spirochaeta thermophila: position and implications for the systematics of the order Spirochaetales. Syst. Appl. Microbiol 16:224–226
    [Google Scholar]
  34. Rainey F. A., Fritze D., Stackebrandt E. 1994; The phylogenetic diversity of thermophilic members of the genus Bacillus as revealed by 16S rDNA analysis. FEMS Microbiol. Lett 115:205–212
    [Google Scholar]
  35. Rainey F. A., Stackebrandt E. 1993; 16S rDNA analysis reveals phylogenetic diversity among the polysaccharolytic clostridia. FEMS Microbiol. Lett 113:125–128
    [Google Scholar]
  36. Shida O., Takagi H., Kadowaki K., Yano H., Abe M., Udaka S., Komagata K. 1994; Bacillus aneurinolyticus sp. nov., nom. rev. Int. J. Syst. Bacteriol 44:143–150
    [Google Scholar]
  37. Sleytr U., Adam H., Klaushofer H. 1969; Die Feinstruktur der Zellwand und Cytoplasmamembran von Clostridium nigrificans, dargestellt mit Hilfe der Gefrieratzund Ultradunnschnittechnik. Arch. Mikrobiol 66:4058
    [Google Scholar]
  38. SJeytr U. B. 1978; Regular arrays of macromolecules on bacterial cell walls: structure, chemistry, assembly, and function. Int Rev. Cytol 53:1–64
    [Google Scholar]
  39. Sleytr U. B., Messner P., Pum D., Sara M. 1993; Crystalline bacterial cell surface layers. Mol. Microbiol 10:911–916
    [Google Scholar]
  40. Sleytr U. B., Messner P., Pum D., Sara M.ed 1996 Crystalline bacterial cell surface proteins R. G. Landes Co.; Austin, Tex:
    [Google Scholar]
  41. Sneath P. H. A. 1986 Endospore-forming Gram-positive rods and cocci. 1104–1207 Sneath P. H. A., Mair N. S., Sharpe M. E., Holt J. G.ed Bergey’s manual of systematic bacteriology 2 Williams & Wilkins; Baltimore:
    [Google Scholar]
  42. Stackebrandt E., Goebel B. M. 1994; A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst Bacteriol 44:846–849
    [Google Scholar]
  43. Takagi H., Shida O., Kadowaki K., Komagata K., Udaka S. 1993; Characterization of Bacillus brevis with descriptions of Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov. Int. J. Syst. Bacteriol 43:221–231
    [Google Scholar]
  44. Tindall B. J. 1990; Lipid composition of Halobacterium lacusprofundi. FEMS Microbiol. Lett 66:199–202
    [Google Scholar]
/content/journal/ijsem/10.1099/00207713-46-2-532
Loading
/content/journal/ijsem/10.1099/00207713-46-2-532
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error