Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1969 Oct;100(1):254–259. doi: 10.1128/jb.100.1.254-259.1969

Specificity of a Bacteriolytic Enzyme from Pseudomonas aeruginosa1

Marvin Lache 1,2,2, Walter R Hearn 1,2, Judith W Zyskind 1,2,2, Donald J Tipper 1,2, Jack L Strominger 1,2,3
PMCID: PMC315386  PMID: 4981057

Abstract

A bacteriolytic enzyme isolated from shake-flask cultures of Pseudomonas aeruginosa and capable of lysing cells of Staphylococcus aureus was purified approximately 500-fold by passage through diethylaminoethyl cellulose and chromatography on carboxymethyl-cellulose. The purified enzyme was shown to act as an endopeptidase, cleaving the pentaglycine cross-bridges of the cell wall peptidoglycan at d-alanyl-glycine and glycyl-glycine linkages with the release of di−, tri−, and tetraglycine fragments. Release of NH2-alanine indicated weak N-acetylmuramyl-l-alanine amidase activity, but most of the residual peptide remained attached to the glycan. No hydrolysis of the glycan occurred. The lytic spectrum of the enzyme toward a variety of other cell walls of known peptidoglycan composition indicated relatively high specificity for peptidoglycans with polyglycine bridges.

Full text

PDF
255

Selected References

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

  1. BROWDER H. P., ZYGMUNT W. A., YOUNG J. R., TAVORMINA P. A. LYSOSTAPHIN: ENZYMATIC MODE OF ACTION. Biochem Biophys Res Commun. 1965 Apr 23;19:383–389. doi: 10.1016/0006-291x(65)90473-0. [DOI] [PubMed] [Google Scholar]
  2. Burke M. E., Pattee P. A. Purification and characterization of a staphylolytic enzyme from Pseudomonas aeruginosa. J Bacteriol. 1967 Mar;93(3):860–865. doi: 10.1128/jb.93.3.860-865.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. GHUYSEN J. M., STROMINGER J. L. STRUCTURE OF THE CELL WALL OF STAPHYLOCOCCUS AUREUS, STRAIN COPENHAGEN. I. PREPARATION OF FRAGMENTS BY ENZYMATIC HYDROLYSIS. Biochemistry. 1963 Sep-Oct;2:1110–1119. doi: 10.1021/bi00905a035. [DOI] [PubMed] [Google Scholar]
  4. GHUYSEN J. M., STROMINGER J. L. STRUCTURE OF THE CELL WALL OF STAPHYLOCOCCUS AUREUS, STRAIN COPENHAGEN. II. SEPARATION AND STRUCTURE OF DISACCHARIDES. Biochemistry. 1963 Sep-Oct;2:1119–1125. doi: 10.1021/bi00905a036. [DOI] [PubMed] [Google Scholar]
  5. GHUYSEN J. M., TIPPER D. J., STROMINGER J. L. STRUCTURE OF THE CELL WALL OF STAPHYLOCOCCUS AUREUS, STRAIN COPENHAGEN. IV. THE TEICHOIC ACID-GLYCOPEPTIDE COMPLEX. Biochemistry. 1965 Mar;4:474–485. doi: 10.1021/bi00879a016. [DOI] [PubMed] [Google Scholar]
  6. Ghuysen J. M., Bricas E., Lache M., Leyh-Bouille M. Structure of the cell walls of Micrococcus lysodeikticus. 3. Isolation of a new peptide dimer, N-alpha-[L-alanyl-gamma-(alpha-D-glutamylglycine)]-L-lysyl-D-alanyl-N-alpha-[L-alanyl-gamma-(alpha-D-glutamylglycine)]-L-lysyl-D-alanine. Biochemistry. 1968 Apr;7(4):1450–1460. doi: 10.1021/bi00844a030. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. MITCHELL P., MOYLE J. Autolytic release and osmotic properties of protoplasts from Staphylococcus aureus. J Gen Microbiol. 1957 Feb;16(1):184–194. doi: 10.1099/00221287-16-1-184. [DOI] [PubMed] [Google Scholar]
  9. PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
  10. PATTEE P. A., BALDWIN J. N. Transduction of resistance to chlortetracycline and novobiocin in Staphylococcus aureus. J Bacteriol. 1961 Dec;82:875–881. doi: 10.1128/jb.82.6.875-881.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Petit J. F., Munoz E., Ghuysen J. M. Peptide cross-links in bacterial cell wall peptidoglycans studied with specific endopeptidases from Streptomyces albus G. Biochemistry. 1966 Aug;5(8):2764–2776. doi: 10.1021/bi00872a037. [DOI] [PubMed] [Google Scholar]
  12. Strominger J. L., Ghuysen J. M. Mechanisms of enzymatic bacteriaolysis. Cell walls of bacteri are solubilized by action of either specific carbohydrases or specific peptidases. Science. 1967 Apr 14;156(3772):213–221. doi: 10.1126/science.156.3772.213. [DOI] [PubMed] [Google Scholar]
  13. TROLLER J. A., FRAZIER W. C. Repression of Staphylococcus aureus by food bacteria. II. Causes of inhibition. Appl Microbiol. 1963 Mar;11:163–165. doi: 10.1128/am.11.2.163-165.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tipper D. J., Strominger J. L., Ensign J. C. Structure of the cell wall of Staphylococcus aureus, strain Copenhagen. VII. Mode of action of the bacteriolytic peptidase from Myxobacter and the isolation of intact cell wall polysaccharides. Biochemistry. 1967 Mar;6(3):906–920. doi: 10.1021/bi00855a035. [DOI] [PubMed] [Google Scholar]
  15. ZYSKIND J. W., PATTEE P. A., LACHE M. STAPHYLOLYTIC SUBSTANCE FROM A SPECIES OF PSEUDOMONAS. Science. 1965 Mar 19;147(3664):1458–1459. doi: 10.1126/science.147.3664.1458. [DOI] [PubMed] [Google Scholar]

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

RESOURCES