Reference : Isolation of the membrane-bound 26 000-Mr penicillin-binding protein of Streptomyces ...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/83022
Isolation of the membrane-bound 26 000-Mr penicillin-binding protein of Streptomyces strain K15 in the form of a penicillin-sensitive D-alanyl-D-alanine-cleaving transpeptidase
English
Nguyen-Distèche, Martine[Université de Liège - ULiège > Faculté de Médecine, Institut de Chimie > Service de Microbiologie > >]
Leyh-Bouille, Mélina[Université de Liège - ULiège > Faculté de Médecine, Institut de Chimie > Service de Microbiologie > > >]
Ghuysen, Jean-Marie[Université de Liège - ULiège > Faculté de Médecine, Institut de Chimie > Service de Microbiologie > > >]
[en] 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.
Fonds de la Recherche Scientifique Médicale - FRSM ; National Institutes of Health - NIH