Abstract
A cell wall lytic enzyme has been demonstrated to be a component of the membrane of Bacillus licheniformis NCTC 6346 and an l-form derived from it. The lytic enzyme, characterized as an N-acetylmuramyl-l-alanine amidase, is solubilized from membranes by nonionic detergents. Ionic detergents inactivate the enzyme. In the bacterium the specific activities of amidase and d-alanine carboxypeptidase in mesosomes are approximately 65% of those in membranes. Selective transfer of lytic enzyme from nongrowing L-forms, L-form membranes, and protoplasts to added walls occurred after mixing, and 31 to 77% of the enzyme lost from L-form membranes was recovered on the walls. Membranes isolated from L-forms growing in the presence of added walls contained as little as 13% of the amidase found in membranes of a control culture. These results have been interpreted as showing that in vivo the amidase is “bound” to the surface of the bacterial cell membrane in such a location that it can be readily accessible to the cell wall.
Full text
PDF![878](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/34a223e48b22/jbacter00362-0100.png)
![879](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/c6f07d20b870/jbacter00362-0101.png)
![880](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/227356450eee/jbacter00362-0102.png)
![881](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/96e91e82e2d8/jbacter00362-0103.png)
![882](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/feacab863d48/jbacter00362-0104.png)
![883](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/72b0f0aa9f0e/jbacter00362-0105.png)
![884](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/400bf0d913fc/jbacter00362-0106.png)
![885](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/50fcb48d4813/jbacter00362-0107.png)
![886](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/6532bfd86e3a/jbacter00362-0108.png)
![887](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/c950e111ee57/jbacter00362-0109.png)
![888](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/247506/3919377cb48c/jbacter00362-0110.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ARRIGONI O., SINGER T. P. Limitations of the phenazine methosulphate assay for succinic and related dehydrogenases. Nature. 1962 Mar 31;193:1256–1258. doi: 10.1038/1931256a0. [DOI] [PubMed] [Google Scholar]
- Arvidson S., Holme T., Wadström T. Formation of bacteriolytic enzymes in batch and continuous culture of Staphylococcus aureus. J Bacteriol. 1970 Oct;104(1):227–233. doi: 10.1128/jb.104.1.227-233.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BRUMFITT W., WARDLAW A. C., PARK J. T. Development of lysozyme-resistance in Micrococcus lysodiekticus and its association with an increased O-acetyl content of the cell wall. Nature. 1958 Jun 28;181(4626):1783–1784. doi: 10.1038/1811783a0. [DOI] [PubMed] [Google Scholar]
- Blumberg P. M., Strominger J. L. Inactivation of D-alanine carboxypeptidase by penicillins and cephalosporins is not lethal in Bacillus subtilis. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2814–2817. doi: 10.1073/pnas.68.11.2814. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown W. C., Fraser D. K., Young F. E. Problems in purification of a Bacillus subtilis autolytic enzyme caused by association with teichoic acid. Biochim Biophys Acta. 1970 Feb 11;198(2):308–315. doi: 10.1016/0005-2744(70)90063-x. [DOI] [PubMed] [Google Scholar]
- Chaloupka J., Krecková P. Turnover of mucopeptide during the life cycle of Bacillus megaterium. Folia Microbiol (Praha) 1971;16(5):372–382. doi: 10.1007/BF02875757. [DOI] [PubMed] [Google Scholar]
- Chatterjee A. N., Mirelman D., Singer H. J., Park J. T. Properties of a novel pleiotropic bacteriophage-resistant mutant of Staphylococcus aureus H. J Bacteriol. 1969 Nov;100(2):846–853. doi: 10.1128/jb.100.2.846-853.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clive D., Landman O. E. Reversion of Bacillus subtilis protoplasts to the bacillary form induced by exogenous cell wall, bacteria and by growth in membrane filters. J Gen Microbiol. 1970 May;61(2):233–243. doi: 10.1099/00221287-61-2-233. [DOI] [PubMed] [Google Scholar]
- Ellwood D. C., Tempest D. W. Control of teichoic acid and teichuronic acid biosyntheses in chemostat cultures of Bacillus subtilis var. niger. Biochem J. 1969 Jan;111(1):1–5. doi: 10.1042/bj1110001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fan D. P. Cell wall binding properties of the Bacillus subtilis autolysin(s). J Bacteriol. 1970 Aug;103(2):488–493. doi: 10.1128/jb.103.2.488-493.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferrandes B., Frehel C., Chaix P. Fractionment et purification des systèmes membranaires cytoplasmiques et mésosomiquees de lbacillus subtilis. Etude de quelques-unes de leurs propríetés oxydo-réductricwa associées à la chaine respiratoire. Biochim Biophys Acta. 1970 Dec 8;223(2):292–308. doi: 10.1016/0005-2728(70)90186-6. [DOI] [PubMed] [Google Scholar]
- Fodor M., Rogers H. J. Antagonism between vegetative cells and L-forms of Bacillus licheniformis strain 6346. Nature. 1966 Aug 6;211(5049):658–659. doi: 10.1038/211658a0. [DOI] [PubMed] [Google Scholar]
- Forsberg C., Rogers H. J. Autolytic enzymes in growth of bacteria. Nature. 1971 Jan 22;229(5282):272–273. doi: 10.1038/229272a0. [DOI] [PubMed] [Google Scholar]
- Hayano K., Fukui S. Purification and properties of 3-ketosucrose-forming enzyme from the cells of Agrobacterium tumefaciens. J Biol Chem. 1967 Aug 25;242(16):3655–3672. [PubMed] [Google Scholar]
- Huff E., Silverman C. S. Lysis of Staphylococcus aureus cell walls by a soluble staphylococcal enzyme. J Bacteriol. 1968 Jan;95(1):99–106. doi: 10.1128/jb.95.1.99-106.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Lawrence P. J., Strominger J. L. Biosynthesis of the peptidoglycan of bacterial cell walls. XVI. The reversible fixation of radioactive penicillin G to the D-alanine carboxypeptidase of Bacillus subtilis. J Biol Chem. 1970 Jul 25;245(14):3660–3666. [PubMed] [Google Scholar]
- MASTER R. W. POSSIBLE SYNTHESIS OF POLYRIBONUCLEOTIDES OF KNOWN BASE-TRIPLET SEQUENCES. Nature. 1965 Apr 3;206:93–93. doi: 10.1038/206093b0. [DOI] [PubMed] [Google Scholar]
- MILLER G. L., GOLDER R. H. Buffers of pH 2 to 12 for use in electrophoresis. Arch Biochem. 1950 Dec;29(2):420–423. [PubMed] [Google Scholar]
- Mauck J., Chan L., Glaser L. Turnover of the cell wall of Gram-positive bacteria. J Biol Chem. 1971 Mar 25;246(6):1820–1827. [PubMed] [Google Scholar]
- Pooley H. M., Porres-Juan J. M., Shockman G. D. Dissociation of an autolytic enzyme-cell wall complex by treatment with unusually high concentrations of salt. Biochem Biophys Res Commun. 1970 Mar 27;38(6):1134–1140. doi: 10.1016/0006-291x(70)90357-8. [DOI] [PubMed] [Google Scholar]
- Pooley H. M., Shockman G. D. Relationship between the latent form and the active form of the autolytic enzyme of Streptococcus faecalis. J Bacteriol. 1969 Nov;100(2):617–624. doi: 10.1128/jb.100.2.617-624.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RICHMOND M. H. Formation of a lytic enzyme by a strain of Bacillus subtilis. Biochim Biophys Acta. 1959 May;33(1):78–92. doi: 10.1016/0006-3002(59)90500-1. [DOI] [PubMed] [Google Scholar]
- Reaveley D. A., Rogers H. J. Some enzymic activities and chemical properties of the mesosomes and cytoplasmic membranes of Bacillus licheniformis 6346. Biochem J. 1969 Jun;113(1):67–79. doi: 10.1042/bj1130067. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rogers H. J. Bacterial growth and the cell envelope. Bacteriol Rev. 1970 Jun;34(2):194–214. doi: 10.1128/br.34.2.194-214.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- SHOCKMAN G. D. Alkaline labile D-alanine in cell walls. Nature. 1963 Jun 8;198:997–999. doi: 10.1038/198997a0. [DOI] [PubMed] [Google Scholar]
- Shockman G. D., Pooley H. M., Thompson J. S. Autolytic enzyme system of Streptococcus faecalis. 3. Localization of the autolysin at the sites of cell wall synthesis. J Bacteriol. 1967 Nov;94(5):1525–1530. doi: 10.1128/jb.94.5.1525-1530.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shockman G. D., Thompson J. S., Conover M. J. The autolytic enzyme system of Streptococcus faecalis. II. Partial characterization of the autolysin and its substrate. Biochemistry. 1967 Apr;6(4):1054–1065. doi: 10.1021/bi00856a014. [DOI] [PubMed] [Google Scholar]
- Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takebe I., Singer H. J., Wise E. M., Jr, Park J. T. Staphylococcus aureus H autolytic activity: general properties. J Bacteriol. 1970 Apr;102(1):14–19. doi: 10.1128/jb.102.1.14-19.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson J. S., Shockman G. D. A modification of the Park and Johnson reducing sugar determination suitable for the assay of insoluble materials: its application to bacterial cell walls. Anal Biochem. 1968 Feb;22(2):260–268. doi: 10.1016/0003-2697(68)90315-1. [DOI] [PubMed] [Google Scholar]
- Van Heijenoort Y., Van Heijenoort J. Study of the N-acetylmuramyl-L-alanine amidase activity in Escherichia coli. FEBS Lett. 1971 Jun 10;15(2):137–141. doi: 10.1016/0014-5793(71)80041-8. [DOI] [PubMed] [Google Scholar]
- WORK E. Reaction of ninhydrin in acid solution with straight-chain amino acids containing two amino groups and its application to the estimation of alpha epsilon-diaminopimelic acid. Biochem J. 1957 Nov;67(3):416–423. doi: 10.1042/bj0670416. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young F. E. Autolytic enzyme associated with cell walls of Bacillus subtilis. J Biol Chem. 1966 Aug 10;241(15):3462–3467. [PubMed] [Google Scholar]