Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1975 Nov;124(2):623–632. doi: 10.1128/jb.124.2.623-632.1975

Formation of cell wall polymers by reverting protoplasts of Bacillus licheniformis.

T S Elliott, J B Ward, H J Rogers
PMCID: PMC235948  PMID: 1184574

Abstract

The biosynthesis of peptidoglycan and teichoic acid by reverting protoplasts of Bacillus licheniformis 6346 His-, in cubated at 35 C on medium containing 2.5% agar, is detectable after 40 min. The amount of N-acetyl-[1-14C]glucosamine incorporated into peptidoglycan and teichoic acid on continued incubation doubles at the same rate as the incorporation of [3H]tryptophan into protein. At the early stages of reversion the average glycan chain length, measured by the ratio of free reducing groups of muramic acid and glucosamine to total muramic acid present, is very short. As reversion proceeds, the average chain length increases to a value similar to the found in the wall of the parent bacillus. The extent of cross-linkage found in the peptide side chains of the peptidoglycan also increases as reversion proceeds. At the completion of reversion the wall material synthesized has similar characteristics to those of the walls of the parent bacilli, containing peptidoglycan and teichoic and teichuronic acids in about the same proportions. Soluble peptidoglycan can be isolated from the reversion medium, amounting to 30% of the total formed after 3 h of incubation and 8% after 12 h. This amount was reduced by the presence in the medium of the walls of an autolysin-deficient mutant; they were not formed at all by reverting protoplasts of the autolysin-deficient mutant itself. Analysis of the soluble material provided additional evidence for their being autolytic products rather than small unchanged molecules. When protoplasts were incubated on medium containing only 0.8% agar, 53 to 67% of the peptidoglycan formed after 3 h of incubation was soluble, and 21% after 12 h. Fibers that appeared to be sheared from the protoplasts at intermediate stages of reversion on medium containing 2.5% agar were similar in composition to the bacillary walls.

Full text

PDF
626

Selected References

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

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Berberich R., Kaback M., Freese E. D-amino acids as inducers of L-alanine dehydrogenase in Bacillus subtilis. J Biol Chem. 1968 Mar 10;243(5):1006–1011. [PubMed] [Google Scholar]
  3. Elliott T. S., Ward J. B., Wyrick P. B., Rogers H. J. Ultrastructural study of the reversion of protoplasts of Bacillus licheniformis to bacilli. J Bacteriol. 1975 Nov;124(2):905–917. doi: 10.1128/jb.124.2.905-917.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fordham W. D., Gilvarg C. Kinetics of cross-linking of peptidoglycan in Bacillus megaterium. J Biol Chem. 1974 Apr 25;249(8):2478–2482. [PubMed] [Google Scholar]
  5. Forsberg C. W., Ward J. B. N-acetylmuramyl-L-alanine amidase of Bacillus licheniformis and its L-form. J Bacteriol. 1972 Jun;110(3):878–888. doi: 10.1128/jb.110.3.878-888.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Forsberg C. W., Wyrick P. B., Ward J. B., Rogers H. J. Effect of phosphate limitation on the morphology and wall composition of Bacillus licheniformis and its phosphoglucomutase-deficient mutants. J Bacteriol. 1973 Feb;113(2):969–984. doi: 10.1128/jb.113.2.969-984.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HANES C. S., ISHERWOOD F. A. Separation of the phosphoric esters on the filter paper chromatogram. Nature. 1949 Dec 31;164(4183):1107-12, illust. doi: 10.1038/1641107a0. [DOI] [PubMed] [Google Scholar]
  8. Hughes R. C. Autolysis of isolated cell walls of Bacillus licheniformis N.C.T.C. 6346 and Bacillus subtilis Marburg Strain 168. Separation of the products and characterization of the mucopeptide fragments. Biochem J. 1970 Oct;119(5):849–860. doi: 10.1042/bj1190849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hughes R. C., Pavlik J. G., Rogers H. J., Tanner P. J. Organization of polymers in the cell walls of some bacilli. Nature. 1968 Aug 10;219(5154):642–644. doi: 10.1038/219642a0. [DOI] [PubMed] [Google Scholar]
  10. Hughes R. C., Tanner P. J. The action of dilute alkali on some bacterial cell walls. Biochem Biophys Res Commun. 1968 Oct 10;33(1):22–28. doi: 10.1016/0006-291x(68)90248-9. [DOI] [PubMed] [Google Scholar]
  11. King J. R., Gooder H. Reversion to the streptococcal state of enterococcal protoplasts, spheroplasts, and L-forms. J Bacteriol. 1970 Sep;103(3):692–696. doi: 10.1128/jb.103.3.692-696.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LEVVY G. A., MCALLAN A. The N-acetylation and estimation of hexosamines. Biochem J. 1959 Sep;73:127–132. doi: 10.1042/bj0730127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Landman O. E., Forman A. Gelatin-induced reversion of protoplasts of Bacillus subtilis to the bacillary form: biosynthesis of macromolecules and wall during successive steps. J Bacteriol. 1969 Aug;99(2):576–589. doi: 10.1128/jb.99.2.576-589.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Landman O. E., Ryter A., Fréhel C. Gelatin-induced reversion of protoplasts of Bacillus subtilis to the bacillary form: electron-microscopic and physical study. J Bacteriol. 1968 Dec;96(6):2154–2170. doi: 10.1128/jb.96.6.2154-2170.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rogers H. J., Forsberg C. W. Role of autolysins in the killing of bacteria by some bactericidal antibiotics. J Bacteriol. 1971 Dec;108(3):1235–1243. doi: 10.1128/jb.108.3.1235-1243.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rogers H. J., Thurman P. F., Taylor C., Reeve J. N. Mucopeptide synthesis by rod mutants of Bacillus subtilis. J Gen Microbiol. 1974 Dec;85(2):335–349. doi: 10.1099/00221287-85-2-335. [DOI] [PubMed] [Google Scholar]
  18. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  19. Ward J. B. The chain length of the glycans in bacterial cell walls. Biochem J. 1973 Jun;133(2):395–398. doi: 10.1042/bj1330395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ward J. B. The synthesis of peptidoglycan in an autolysin-deficient mutant of Bacillus licheniformis N.C.T.C. 6346 and the effect of beta-lactam antibiotics, bacitracin and vancomycin. Biochem J. 1974 Jul;141(1):227–241. doi: 10.1042/bj1410227. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES