title: Overproduction and purification of the Aeromonas hydrophila CphA metallo-beta-lactamase expressed in Escherichia coli.
abstract: The Aeromonas hydrophila CphA metallo-beta-lactamase was overexpressed in a soluble secreted form in Escherichia coli using a T7 RNA polymerase-based expression system, and a simple protocol based on a single cation-exchange chromatographic step was developed, which is suitable for rapid purification of the overexpressed enzyme from E. coli lysates. A yield of up to 30 micrograms of purified enzyme per milliliter of culture was obtained. The purified enzyme preparation showed properties identical to those previously reported in the literature.
authors.affiliation: Dipartimento di Scienze e Tecnologie Biomediche e di Biometria, Università dell'Aquila, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Hernandez Villadares
authors.forename: M
authors.initials: M
authors.lastname: Galleni
authors.forename: J M
authors.initials: JM
authors.lastname: Frère
authors.forename: A
authors.initials: A
authors.lastname: Felici
authors.forename: M
authors.initials: M
authors.lastname: Perilli
authors.forename: N
authors.initials: N
authors.lastname: Franceschini
authors.forename: G M
authors.initials: GM
authors.lastname: Rossolini
authors.forename: A
authors.initials: A
authors.lastname: Oratore
authors.forename: G
authors.initials: G
authors.lastname: Amicosante
mesh_terms: D016980:Aeromonas hydrophila / Q000201:enzymology* / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005822:Genetic Vectors; D006868:Hydrolysis; D015378:Imipenem / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; D011994:Recombinant Proteins; D015378:Imipenem; D001618:beta-Lactamases; C082063:cphA protein, Aeromonas hydrophila