title: Secretion by overexpression and purification of the water-soluble Streptomyces K15 DD-transpeptidase/penicillin-binding protein.
abstract: Though synthesized with a cleavable signal peptide and devoid of membrane anchors, the 262-amino-acid-residue Streptomyces K15 DD-transpeptidase/penicillin-binding protein is membrane-bound. Overexpression in Streptomyces lividans resulted in the export of an appreciable amount of the synthesized protein (4 mg/litre of culture supernatant). The water-soluble enzyme was purified close to protein homogeneity with a yield of 75%. It requires the presence of 0.5 M-NaCl to remain soluble. It is indistinguishable from the detergent-extract wild-type enzyme with respect to molecular mass, thermostability, transpeptidase activity and penicillin-binding capacity.
authors.affiliation: Centre d'Ingénierie des Protéines, Université de Liège, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Palomeque-Messia
authors.forename: V
authors.initials: V
authors.lastname: Quittre
authors.forename: M
authors.initials: M
authors.lastname: Leyh-Bouille
authors.forename: M
authors.initials: M
authors.lastname: Nguyen-Distèche
authors.forename: C J
authors.initials: CJ
authors.lastname: Gershater
authors.forename: I K
authors.initials: IK
authors.lastname: Dacey
authors.forename: J
authors.initials: J
authors.lastname: Dusart
authors.forename: J
authors.initials: J
authors.lastname: Van Beeumen
authors.forename: J M
authors.initials: JM
authors.lastname: Ghuysen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D004795:Enzyme Stability; D015870:Gene Expression; D006602:Hexosyltransferases; D006358:Hot Temperature; D008969:Molecular Sequence Data; D008970:Molecular Weight; D002267:Muramoylpentapeptide Carboxypeptidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D046915:Penicillin-Binding Proteins; D010406:Penicillins / Q000378:metabolism; D010458:Peptidyl Transferases; D010957:Plasmids; D012965:Sodium Chloride; D012995:Solubility; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014169:Transformation, Bacterial
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D046915:Penicillin-Binding Proteins; D010406:Penicillins; D012965:Sodium Chloride; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase