Abstract
The membrane-bound, 26 000-Mr penicillin-binding protein of Streptomyces K15 has been isolated in the form of an effective, penicillin-sensitive D-alanyl-D-alanine-cleaving peptidase exhibiting high transpeptidase activity (greater than 95%) and very low carboxy-peptidase activity (less than 5%). The penicillin-binding protein/transpeptidase can be extracted directly from the mycelium with N-cetyl-NNN-trimethylammonium bromide (Cetavlon) and subsequently obtained at 90% purity and with an 8000-fold specific enrichment (when compared with the activity of the isolated membranes) by a two-step procedure involving Sephadex filtration and affinity chromatography on ampicillin-linked CH Sepharose 4B in the presence of detergent. At saturating concentrations of the co-substrates diacetyl-L-Lys-D-Ala-D-Ala and Gly-Gly, the catalytic-centre activity is about 0.3 s-1.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Coyette J., Ghuysen J. M., Fontana R. Solubilization and isolation of the membrane-bound DD-carboxypeptidase of Streptococcus faecalis ATCC9790. Properties of the purified enzyme. Eur J Biochem. 1978 Jul 17;88(1):297–305. doi: 10.1111/j.1432-1033.1978.tb12450.x. [DOI] [PubMed] [Google Scholar]
- Dusart J., Leyh-Bouille M., Ghuysen J. M. The peptidoglycan crosslinking enzyme system in Streptomyces strains R61, K15 and rimosus. Kinetic coefficients involved in the interactions of the membrane-bound transpeptidase with peptide substrates and beta-lactam antibiotics. Eur J Biochem. 1977 Nov 15;81(1):33–44. doi: 10.1111/j.1432-1033.1977.tb11924.x. [DOI] [PubMed] [Google Scholar]
- Frère J. M., Ghuysen J. M., Perkins H. R., Nieto M. Molecular weight and amino acid composition of the exocellular DD-carboxypeptidase-transpeptidase of Streptomyces R61. Biochem J. 1973 Nov;135(3):463–468. doi: 10.1042/bj1350463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leyh-Bouille M., Dusart J., Nguyen-Distèche M., Ghuysen J. M., Reynolds P. E., Perkins H. R. The peptidoglycan crosslinking enzyme system in Streptomyces strains R61, K15 and rimosus. Eur J Biochem. 1977 Nov 15;81(1):19–28. doi: 10.1111/j.1432-1033.1977.tb11922.x. [DOI] [PubMed] [Google Scholar]
- Leyh-Bouille M., Nguyen-Distèche M., Ghuysen J. M. On the DD-carboxypeptidase enzyme system of Streptomyces strain K15. Eur J Biochem. 1981 Apr;115(3):579–584. doi: 10.1111/j.1432-1033.1981.tb06242.x. [DOI] [PubMed] [Google Scholar]
- Marquet A., Dusart J., Ghuysen J. M., Perkins H. R. Membrane-bound transpeptidase and penicillin binding sites in Streptomyces strain R61. Eur J Biochem. 1974 Aug 1;46(3):515–523. doi: 10.1111/j.1432-1033.1974.tb03645.x. [DOI] [PubMed] [Google Scholar]
- Matsuzawa H., Datta P., Matsuhashi M. Behavior of penicillin-binding proteins in Escherichia coli upon heat and detergent treatments and partial purification of penicillin-binding proteins 1A and 1B. J Bacteriol. 1979 Jun;138(3):1029–1032. doi: 10.1128/jb.138.3.1029-1032.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moore B. A., Brammer K. W. 6 beta-Iodopenicillanic acid (UI-38,006), a beta-lactamase inhibitor that extends the antibacterial spectrum of beta-lactam compounds: initial bacteriological characterization. Antimicrob Agents Chemother. 1981 Sep;20(3):327–331. doi: 10.1128/aac.20.3.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spratt B. G. Biochemical and genetical approaches to the mechanism of action of penicillin. Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):273–283. doi: 10.1098/rstb.1980.0045. [DOI] [PubMed] [Google Scholar]