Protoplast and Cytoplasmic Membrane Preparations from and Free

Abstract

Protoplasts were prepared from and some serotypes by use of lysozyme (EC 3.2.1.17) under particular conditions: cells had to be grown in -threonine (20 m) and harvested in early exponential phase. The efficiency of protoplast formation was enhanced by two additional steps: plasmolysis (in 12% PEG), prior to addition of lysozyme, and a swirling phase, after the enzymic action. This procedure allowed us to obtain clean protoplasts, with only 0·5% contamination by bacterial cell walls. Up to 90% protoplast lysis was obtained in 0·5 -NaCl. Cytoplasmic membrane purification was achieved by centrifugation on a glycerol cushion.

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1983-10-01
2024-03-28
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References

  1. Abrams A. 1965; The release of bound adenosine triphosphatase from isolated bacterial membranes and the properties of the solubilized enzyme. Journal of Biological Chemistry 240:3675–3681
    [Google Scholar]
  2. Baginski E.S., Zak B. 1960; Microdetermination of serum phosphate and phospholipids. Clinica chimica acta 5:834–838
    [Google Scholar]
  3. Beighton D., Russell R.R.B., Hayday H. 1981; The isolation and characterization ofStreptococcus mutans serotypeh from dental plaque of monkeys (Macaca fascicularis). . Journal of General Microbiology 124:271–279
    [Google Scholar]
  4. Bleiweis A.S., Zlmmermann L.B. 1961; Formation of two types of osmotically fragile bodies fromStreptococcus faecalis var.liquefaciens. . Canadian Journal of Microbiology 7:363–373
    [Google Scholar]
  5. Bratthall D. 1970; Demonstration of five serological groups of streptococcal strains resemblingStreptococcus mutans. . Odontologisk revy 21:143–152
    [Google Scholar]
  6. Calandra G.B., Nugent K.M., Cole R.M. 1975; Preparation of protoplasts of group H streptococci (Streptococcus sanguis). . Applied Microbiology 29:90–93
    [Google Scholar]
  7. Chassy B.M. 1976; A gentle method for lysis of oral streptococci. Biochemical and Biophysical Research Communications 68:603–608
    [Google Scholar]
  8. Chiu T.H., Emdur L.I., Platt D. 1974; Lipoteichoic acids fromStreptococcus sanguis. . Journal of Bacteriology 118:471–479
    [Google Scholar]
  9. Coleman S.E., Van De Rijn I., Bleiweis A.S. 1970; Lysis of grouped and ungrouped streptococci by lysozyme. Infection and Immunity 2:563–569
    [Google Scholar]
  10. Coykendall A.L. 1974; Four types ofStreptococcus mutans based on their genetic, antigenic and biochemical characteristics. Journal of General Microbiology 83:327–338
    [Google Scholar]
  11. Dulley J.R., Grieve P.A. 1975; A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Analytical Biochemistry 64:136–141
    [Google Scholar]
  12. Eisenberg R.J., Lillmars K. 1975; A method for gentle lysis ofStreptococcus mutans. . Biochemical and Biophysical Research Communications 65:378–384
    [Google Scholar]
  13. Formanek H., Formanek S., Wawra H. 1974; A three dimensional atomic model of the murein layer of bacteria. European Journal of Biochemistry 46:279–294
    [Google Scholar]
  14. Gibbons R.J., De Stoppelaar J.D., Harden L. 1966; Lysozyme insensitivity of bacteria indigenous to the oral cavity of man. Journal of Dental Research 45:877–881
    [Google Scholar]
  15. Goodman H., Pollock J.J., Katona L.I., Iacono V.J., Cho M.I., Thomas E. 1981; Lysis ofStreptococcus mutans by hen egg white lysozyme and inorganic sodium salts. Journal of Bacteriology 146:764–774
    [Google Scholar]
  16. Guggenheim B. 1968; Streptococci of dental plaque. Caries Research 2:147–163
    [Google Scholar]
  17. Hayashi H., Araki Y., Ito E. 1973; Occurrence of glucosamine residues with free amino groups in cell wall peptidoglycan from bacilli as a factor responsible for resistance to lysozyme. Journal of Bacteriology 113:592–598
    [Google Scholar]
  18. Helenius A., Simons K. 1975; Solubilization of membranes by detergents. Biochimica et biophysica acta 415:29–79
    [Google Scholar]
  19. Johnston K.H., Gotschlich E.C. 1974; Isolation and characterization of the outer membrane ofNeisseria gonorrhoeae. . Journal of Bacteriology 119:250–257
    [Google Scholar]
  20. Joseph R., Shockman G.D. 1975; Cellular localization of lipoteichoic acid inStreptococcus faecalis. . Journal of Bacteriology 122:1375–1386
    [Google Scholar]
  21. Kessler R.E., Van De Rijn I. 1979; Quantitative immunoelectrophoretic analysis ofStreptococcus pyogenes membrane. Infection and Immunity 26:892–902
    [Google Scholar]
  22. Krause R.M. 1972; The antigens of group D streptococci. In Streptococci and Streptococcal Diseases pp. 67–74 Edited by Wannamaker L.W., Matsen J.M. New York: Academic Press;
    [Google Scholar]
  23. Markham J.L., Knox K.W., Wicken A.J., Hewett M.J. 1975; Formation of extracellular lipoteichoic acid by oral streptococci and lactobacilli. Infection and Immunity 12:378–386
    [Google Scholar]
  24. Moldow C., Robertson J., Rothfield L. 1972; Purification of bacterial membrane proteins. The use of guanidium thiocyanate and urea. Journal of Membrane Biology 10:137–152
    [Google Scholar]
  25. Morris D.L. 1948; Quantitative determination of carbohydrates with Dreywood’s anthrone reagent. Science 107:254–256
    [Google Scholar]
  26. Osborn M.J., Gander J.E., Parisi E., Carson J. 1972; Mechanism of assembly of the outer membrane ofSalmonella typhimurium. . Journal of Biological Chemistry 247:3962–3972
    [Google Scholar]
  27. Owen P., Freer J.H. 1972; Isolation and properties of mesosomal membrane fractions fromMicrococcus lysodeikticus. . Biochemical Journal 129:907–917
    [Google Scholar]
  28. Perch B., Kjems E., Ravn T. 1974; Biochemical and serological properties ofStreptococcus mutansfrom various human and animal sources. Acta pathologica et microbiologica scandinavica B, 82:357–370
    [Google Scholar]
  29. Pollock J.J., Iacono V.J., Goodman Bicker H., Mackay B.J., Katona L.I., Taichman L.B., Thomas E. 1976; The binding, aggregation and lytic properties of lysozyme. In Proceedings, Microbial Aspects of Dental Caries 2: pp. 325–352 Edited by Stiles H.M., Loesche W.J., O’Brien T. C. Special supplement toMicrobiology Abstracts. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  30. Russell M.W., Lehner T. 1978; Characterization of antigens extracted from cells and culture fluids ofStreptococcus mutans serotypec. . Archives of Oral Biology 23:7–15
    [Google Scholar]
  31. Russell R.R.B. 1979; Wall-associated protein antigens ofStreptococcus mutans. . Journal of General Microbiology 114:109–115
    [Google Scholar]
  32. Salton M.R.J. 1974; Membrane associated enzymes in bacteria. Advances in Microbial Physiology 11:213–282
    [Google Scholar]
  33. Schöller M., Klein J.P., Frank R.M. 1981; Common antigens of streptococcal and non-streptococcal oral bacteria: immunochemical studies of extracellular and cell-wall associated antigens fromStreptococcus sanguis, Streptococcus mutans, Lactobacillus salivarius andActinomyces viscosus. . Infection and Immunity 31:52–60
    [Google Scholar]
  34. Shklair I.L., Keene H.J. 1976; Biochemical characterization and distribution ofStreptococcus mutans in three diverse populations. In Proceedings, Microbial Aspects of Dental Caries 1 pp. 201–210 Edited by Stiles H.M., Loesche W.J., O’Brien T.C. Special Supplement toMicrobiology Abstracts. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  35. Shockman G.D., Daneo-Moore L., Cornett J.B., Mychajlonka M. 1979; Does penicillin kill bacteria?. Reviews of Infectious Diseases 1:787–796
    [Google Scholar]
  36. Siegel J.L., Hurst S.F., Liberman E.S., Coleman S.E., Bleiweis A.S. 1981; Mutanolysin-induced spheroplasts ofStreptococcus mutans are true protoplasts. Infection and Immunity 31:808–815
    [Google Scholar]
  37. Silvestri L.J., Craig R.A., Ingram L.O., Hoffmann E.M., Bleiweis A.S. 1978; Purification of lipoteichoic acid by using phosphatidyl choline vesicles. Infection and Immunity 22:107–118
    [Google Scholar]
  38. Smith D.G. 1973; The lysozyme sensitivity ofStreptococcus equinus. . Microbios 8:199–207
    [Google Scholar]
  39. Taubman M.A. 1973; Role of immunization in dental disease. In Comparative Immunology of the Oral Cavity pp. 138–158 Edited by Mergenhagen S.E., Sherp H.W. Washington, D.C.: U.S. Government Printing Office;
    [Google Scholar]
  40. Thomas P.S., Farquhar M.N. 1978; Specific measurement of DNA in nuclei and nucleic acids using diaminobenzoic acid. Analytical Biochemistry 89:35–44
    [Google Scholar]
  41. Van De Rijn I., Kessler R.E. 1979; Chemical analysis of changes in membrane composition during growth ofStreptococcus pyogenes. . Infection and Immunity 26:883–891
    [Google Scholar]
  42. Vaught R.M., Bleiweis A.S. 1974; Antigens ofStreptococcus mutans. II. Characterization of an antigen resembling a glycerol teichoic acid in walls of strain BHT. Infection and Immunity 9:60–67
    [Google Scholar]
  43. Without B., Boekhout M. 1978; The effect of osmotic shock on the accessibility of the murein layer of exponentially growingEscherichia coli to lysozyme. Biochimica et biophysica acta 508:296–305
    [Google Scholar]
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