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. 1980 Sep;143(3):1491–1497. doi: 10.1128/jb.143.3.1491-1497.1980

Growth of Streptococcus mutans protoplasts is not inhibited by penicillin.

L C Parks, G D Shockman, M L Higgins
PMCID: PMC294542  PMID: 6997274

Abstract

A method is described in which cells of Streptococcus mutans BHT can be converted to spherical, osmotically fragile protoplasts. Exponential-phase cells were suspended in a solution containing 0.5 M melezitose, and their cell walls were hydrolyzed with mutanolysin (M-1 enzyme). When the resultant protoplasts were incubated in a chemically defined growth medium containing 0.5 M NH4Cl, the protoplast suspensions increased in turbidity, protein, ribonucleic acid, and deoxyribonucleic acid in a balanced fashion. In the presence of benzylpenicillin (5 microgram/ml), balanced growth of protoplasts was indistinguishable from untreated controls. This absence of inhibition of protoplast growth in the presence of benzylpenicillin was apparently not due to inactivation of the antibiotic. When exponential-phase cells of S. mutans BHT were first exposed to 5 microgram of benzyl-penicillin per ml for 1 h and then converted to protoplasts, these protoplasts were also able to grow in chemically defined, osmotically stabilized medium. The ability of wall-free protoplasts to grow and to synthesize ribonucleic acid and protein in the presence of a relatively high concentration of benzylpenicillin contrasts with the previously reported rapid inhibition of ribonucleic acid and protein synthesis in intact streptococci. These data suggest that this secondary inhibition of ribonucleic acid and protein synthesis in whole cells is due to factors involved with the continued assembly of an intact, insoluble cell wall rather than with earlier stages of peptidoglycan synthesis.

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Selected References

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  1. Coleman S. E., van de Rijn I., Bleiweis A. S. Lysis of grouped and ungrouped streptococci by lysozyme. Infect Immun. 1970 Nov;2(5):563–569. doi: 10.1128/iai.2.5.563-569.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Eisenberg R. J., Lillmars K. A method for gentle lysis of Streptococcus sanguis and Streptococcus mutans. Biochem Biophys Res Commun. 1975 Jul 8;65(1):378–384. doi: 10.1016/s0006-291x(75)80104-5. [DOI] [PubMed] [Google Scholar]
  3. Gibbons R. J., de Stoppelaar J. D., Harden L. Lysozyme insensitivity of bacteria indigenous to the oral cavity of man. J Dent Res. 1966 May-Jun;45(3):877–881. doi: 10.1177/00220345660450036201. [DOI] [PubMed] [Google Scholar]
  4. HASH J. H. PURIFICATION AND PROPERTIES OF STAPHYLOLYTIC ENZYMES FROM CHALAROPSIS SP. Arch Biochem Biophys. 1963 Sep;102:379–388. doi: 10.1016/0003-9861(63)90245-5. [DOI] [PubMed] [Google Scholar]
  5. Hancock R., Fitz-James P. C. Some differences in the action of penicillin, bacitracin, and vancomycin on Bacillus megaterium. J Bacteriol. 1964 May;87(5):1044–1050. doi: 10.1128/jb.87.5.1044-1050.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heymann H., Manniello J. M., Barkulis S. S. Structure of streptococcal cell walls. V. Phosphate esters in the walls of group A Streptococcus pyogenes. Biochem Biophys Res Commun. 1967 Feb 21;26(4):486–491. doi: 10.1016/0006-291x(67)90574-8. [DOI] [PubMed] [Google Scholar]
  7. Higgins M. L., Pooley H. M., Shockman G. D. Site of initiation of cellular autolysis in Streptococcus faecalis as seen by electron microscopy. J Bacteriol. 1970 Aug;103(2):504–512. doi: 10.1128/jb.103.2.504-512.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Inoue M., Hamada S., Ooshima T., Kotani S., Kato K. Chemical composition of Streptococcus mutans cell walls and their susceptibility to Flavobacterium L-11 enzyme. Microbiol Immunol. 1979;23(5):319–328. doi: 10.1111/j.1348-0421.1979.tb00469.x. [DOI] [PubMed] [Google Scholar]
  9. Joseph R., Shockman G. D. Autolytic formation of protoplasts (autoplasts) of Streptococcus faecalis 9790: release of cell wall, autolysin, and formation of stable autoplasts. J Bacteriol. 1974 May;118(2):735–746. doi: 10.1128/jb.118.2.735-746.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mattingly S. J., Daneo-Moore L., Shockman G. D. Factors regulating cell wall thickening and intracellular iodophilic polysaccharide storage in Streptococcus mutans. Infect Immun. 1977 Jun;16(3):967–973. doi: 10.1128/iai.16.3.967-973.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mychajlonka M., McDowell T. D., Shockman G. D. Inhibition of peptidoglycan, ribonucleic acid, and protein synthesis in tolerant strains of Streptococcus mutans. Antimicrob Agents Chemother. 1980 Apr;17(4):572–582. doi: 10.1128/aac.17.4.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rosenthal R. S., Jungkind D., Daneo-Moore L., Shockman G. D. Evidence for the synthesis of soluble peptidoglycan fragments by protoplasts of Streptococcus faecalis. J Bacteriol. 1975 Oct;124(1):398–409. doi: 10.1128/jb.124.1.398-409.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roth G. S., Shockman G. D., Daneo-Moore L. Balanced macromolecular biosynthesis in "protoplasts" of Streptococcus faecalis. J Bacteriol. 1971 Mar;105(3):710–717. doi: 10.1128/jb.105.3.710-717.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SHOCKMAN G. D., LAMPEN J. O. Inhibition by antibiotics of the growth of bacterial and yeast protoplasts. J Bacteriol. 1962 Sep;84:508–512. doi: 10.1128/jb.84.3.508-512.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schaeffer P., Cami B., Hotchkiss R. D. Fusion of bacterial protoplasts. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2151–2155. doi: 10.1073/pnas.73.6.2151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shockman G. D., Daneo-Moore L., Cornett J. B., Mychajlonka M. Does penicillin kill bacteria?. Rev Infect Dis. 1979 Sep-Oct;1(5):787–796. doi: 10.1093/clinids/1.5.787. [DOI] [PubMed] [Google Scholar]
  17. Terleckyj B., Willett N. P., Shockman G. D. Growth of several cariogenic strains of oral streptococci in a chemically defined medium. Infect Immun. 1975 Apr;11(4):649–655. doi: 10.1128/iai.11.4.649-655.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tomasz A. From penicillin-binding proteins to the lysis and death of bacteria: a 1979 view. Rev Infect Dis. 1979 May-Jun;1(3):434–467. doi: 10.1093/clinids/1.3.434. [DOI] [PubMed] [Google Scholar]
  19. YUDKIN M. D. THE EFFECT OF PENICILLIN, NOVOBIOCIN, STREPTOMYCIN AND VANCOMYCIN ON MEMBRANE SYNTHESIS BY PROTOPLASTS OF BACILLUS MEGATERIUM. Biochem J. 1963 Nov;89:290–296. doi: 10.1042/bj0890290. [DOI] [PMC free article] [PubMed] [Google Scholar]

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