Archives of Microbiology

, Volume 141, Issue 3, pp 249–254 | Cite as

Induction of autolysis of staphylococci by the basic peptide antibiotics Pep 5 and nisin and their influence on the activity of autolytic enzymes

  • Gabriele Bierbaum
  • Hans-Georg Sahl
Original Papers


Pep 5 and nisin are cationic bactericidal peptides which were shown to induce autolysis in Staphylococcus cohnii 22. In contrast to nisin, Pep 5 induced lysis could be stimulated in the presence of glucose. Addition of lipoteichoic acids (LTA) (d-alanine:phosphorus=0.475:1) inhibited all effects of Pep 5 on susceptible cells in a molar ratio LTA:Pep 5 of 10:1. Treatment of S. cohnii 22 with Pep 5 or nisin for 20 min and subsequent washing with 2.5 M NaCl released autolysin activity. Crude preparations of the hydrolyzing enzymes produced free amino groups as well as polysaccharide fragments from the murein backbone, suggesting the presence of a muramidase or glucosamidase, and endopeptidase or amidase. Both enzyme activities were inhibited by lipoteichoic acid; they could be fully reactivated by addition of Pep 5 in sufficient concentrations. The velocity of hydrolysis was not influenced by nisin, whereas it was doubled in presence of Pep 5. The results are discussed in view of a possible mechanism of induction of lysis by Pep 5 and nisin.

Key words

Staphylococcin-like peptide Pep 5 Nisin Autolytic enzymes of Staphylococcus cohnii 22 Regulation of autolysis Cationic peptides Lipoteichoic acids 



arbitrary unit


carbonylcyanide-m-chlorophenyl hydrazone




casein yeast extract glucose


initial turbidity


lipoteichoic acid




Tryptone Soy Broth


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allsop J, Work E (1963) Cell walls of Propionibacterium species: Fractionation and composition. Biochem J 87:512–519PubMedGoogle Scholar
  2. Aminoff D, Morgan WTJ, Watkins WM (1952) Studies in immunochemistry. 11. The action of dilute alkali on the N-acetylhexosamines and the specific blood-group mucoids. Biochem J 51:379–389PubMedGoogle Scholar
  3. Blümel P, Reinicke B, Lahav M, Giesbrecht P (1983) Cell wall degradation of Staphylococcus aureus by autolysins and lysozyme. In: Hakenbeck R, Höltje JV, Labischinski H (eds) The target of penicillin. The murein sacculus of bacterial cell walls. Architecture and growth. Proceedings International FEMS Symposium Berlin (West). Walter de Gruyter. Berlin New York, pp 323–328Google Scholar
  4. Brandis H, Sahl HG (1984) Staphylococcins and other antibacterial substances produced by staphylococci. In: Meyer W (ed) Staphylokokken und Staphylokokken-Erkrankungen. VEB Gustav Fischer, Jena, pp 173–186Google Scholar
  5. Chen PS jr, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Analyt Chem 28:1756–1758Google Scholar
  6. Cleveland RF, Wicken AJ, Daneo-Moore L, Shockman GD (1976) Inhibition of wall autolysis in Streptococcus faecalis by lipoteichoic acid and lipids. J Bacteriol 126:192–197PubMedGoogle Scholar
  7. Ellison JS, Mattern CFT, Daniel WA (1971) Structural changes in Clostridium botulium type E after treatment with boticin S51. J Bacteriol 108:526–534PubMedGoogle Scholar
  8. Fischer W, Rösel P (1980) The alanine ester substitution of lipoteichoic acid (LTA) in Staphylococcus aureus. FEBS Lett 119:224–226PubMedGoogle Scholar
  9. Fischer W, Koch HU, Rösel P (1980) Alanine ester-containing native lipoteichoic acids do not act as lipoteichoic acid carrier. Isolation, structural and functional characterization. J Biol Chem 255:4557–4562PubMedGoogle Scholar
  10. Fischer W, Rösel P, Koch HU (1981) Effect of alanine ester substitution and other structural features of lipoteichoic acids on their inhibitory activity against autolysins of Staphylococcus aureus. J Bacteriol 146:467–475PubMedGoogle Scholar
  11. Fischer W, Koch HU, Haas R (1983) Improved preparation of lipoteichoic acids. Eur J Biochem 133:523–530PubMedGoogle Scholar
  12. Ghuysen JM, Tipper DJ, Strominger JL (1966) Enzymes that degrade bacterial cell walls. In: Neufield EF, Ginsberg V (eds) Methods in enzymology, vol 8. Academic Press, New York, pp 685–699Google Scholar
  13. Ginsburg I, Lahav M (1983) Lysis and biodegradation of microorganisms in infectious sites may involve cooperation between leukocyte, serum factors and bacterial wall autolysins: a working hypothesis. Eur J Clin Microbiol 2:186–191PubMedGoogle Scholar
  14. Giudicelli S, Tomasz A (1984) Attachment of pneumococcal autolysin to wall teichoic acids, an essential step in enzymatic wall degradation. J Bacteriol 158:1188–1190PubMedGoogle Scholar
  15. Graßl M (1970) d-Alanin. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, 2nd ed, vol II. Verlag Chemie, Weinheim/Bergstraße, pp 1641–1644Google Scholar
  16. Gross E, Morell JL (1971) The structure of nisin. J Am Chem Soc 93:4634–4635PubMedGoogle Scholar
  17. Haas R, Koch HU, Fischer W (1984) Alanyl turnover from lipoteichoic acid to teichoic acid in Staphylococcus aureus. FEMS Microbiol Lett 21:27–31Google Scholar
  18. Herbert D, Phipps PJ, Strange RE (1971) Chemical analysis of microbial cells. In: Norris JR, Ribbons DW (eds) Methods in microbiology, vol 5 B. Academic Press, London New York, pp 210–344Google Scholar
  19. Herbold DR, Glaser L (1975) Interaction of N-acetylmuramic acid l-alanine amidase with cell wall polymers. J Biol Chem 250: 7231–7238PubMedGoogle Scholar
  20. Jolliffe LK, Doyle RJ, Streips UN (1981) The energized membrane and cellular autolysis in Bacillus subtilis. Cell 25:753–763PubMedGoogle Scholar
  21. Krebs KG, Heusser D, Wimmer H (1967) Sprühreagenzien. In: Stahl E (ed) Dünnschichtchromatographie. Ein Laboratoriums-handbuch, 2nd ed. Springer, Berlin Heidelberg New York, pp 813–861Google Scholar
  22. Lahav M, Ginsburg I, Kersten T, Wecke J, Giesbrecht P (1983) Induced autolytic wall processes in heat-inactivated Staphylococcus aureus. In: Hakenbeck R, Höltje JV, Labischinski H (eds) The target of penicillin. The murein sacculus of bacterial cell walls. Architecture and growth. Proceedings International FEMS Symposium Berlin (West). Walter de Gruyter, Berlin New York, pp 335–340Google Scholar
  23. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  24. MacKay BJ, Denepitiya L, Ianoco VJ, Krost SB, Pollock JJ (1984) Growth-inhibitory and bactericidal effects of human parotid salivary histidine-rich polypeptides on Streptococcus mutants. Infect Immun 44:695–701PubMedGoogle Scholar
  25. Ramseier HR (1960) Die Wirkung von Nisin auf Clostridium butyricum Prazm. Arch Mikrobiol 37:57–94PubMedGoogle Scholar
  26. Reisinger P, Seidel H, Tschesche H, Hammes WP (1980) The effect of nisin on murein synthesis. Arch Microbiol 127:187–193PubMedGoogle Scholar
  27. Reissig JL, Strominger JL, Leloir LF (1955) A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem 217:959–966PubMedGoogle Scholar
  28. Sahl HG, Brandis H (1981) Production, purification and chemical properties of an antistaphylococcal agent produced by Staphylococcus epidermidis. J Gen Microbiol 127:377–384PubMedGoogle Scholar
  29. Sahl HG, Brandis H (1982) Mode of action of the staphylococcin-like peptide Pep 5 and culture conditions effecting its activity. Zbl Bakt Hyg, I Abt Orig A 252:166–175Google Scholar
  30. Sahl HG, Brandis H (1983) Efflux of low-M r substances from the cytoplasm of sensitive cells caused by the staphylococcin-like agent Pep 5. FEMS Microbiol Lett 16:75–79CrossRefGoogle Scholar
  31. Schaller K, Höltje JV, Braun V (1982) Colicin M is an inhibitor of murein biosynthesis. J Bacteriol 152:994–1000PubMedGoogle Scholar
  32. Selsted ME, Szklarek D, Lehrer RI (1984) Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes. Infect Immun 45:150–154PubMedGoogle Scholar
  33. Steiner H, Hultmark D, Engström A, Bennich H, Boman HG (1981) Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature 292:246–248PubMedGoogle Scholar
  34. Tipper DJ (1969) Mechanism of autolysis of isolated cell walls of Staphylococcus aureus. J Bacteriol 97:837–847PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Gabriele Bierbaum
    • 1
  • Hans-Georg Sahl
    • 1
  1. 1.Institut für Med. Mikrobiologie und Immunologie der Universität BonnBonnGermany

Personalised recommendations