OVERPRODUCTION AND PURIFICATION OF THE AEROMONAS-HYDROPHILA CPHA METALLO-BETA-LACTAMASE EXPRESSED IN ESCHERICHIA-COLI

Authors
VILLADARES MH GALLENI M FRERE JM FELICI A PERILLI M FRANCESCHINI N ROSSOLINI GM ORATORE A AMICOSANTE G
Citation
Mh. Villadares et al., OVERPRODUCTION AND PURIFICATION OF THE AEROMONAS-HYDROPHILA CPHA METALLO-BETA-LACTAMASE EXPRESSED IN ESCHERICHIA-COLI, Microbial drug resistance, 2(2), 1996, pp. 253-256
Citations number
11
Categorie Soggetti
Microbiology,"Infectious Diseases
Journal title
Microbial drug resistance → ACNP
ISSN journal
10766294
Volume
2
Issue
2
Year of publication
1996
Pages
253 - 256
Database
ISI
SICI code
1076-6294(1996)2:2<253:OAPOTA>2.0.ZU;2-L
Abstract
The Aeromonas hydrophila CphA metallo-beta-lactamase was overexpressed in a soluble secreted form in Escherichia coli using a T7 RNA polymer ase-based expression system, and a simple protocol based on a single c ation-exchange chromatographic step was developed, which is suitable f or rapid purification of the overexpressed enzyme from E, coli lysates . A yield of up to 30 mu g of purified enzyme per milliliter of cultur e was obtained, The purified enzyme preparation showed properties iden tical to those previously reported in the literature.