Overproduction and Purification of the Aeromonas hydrophila CphA Metallo-β-Lactamase Expressed in Escherichia coli
- 1 January 1996
- journal article
- research article
- Published by Mary Ann Liebert Inc in Microbial Drug Resistance
- Vol. 2 (2) , 253-256
- https://doi.org/10.1089/mdr.1996.2.253
Abstract
The Aeromonas hydrophila CphA metallo-β-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 over-expressed enzyme from E. coli lysates. A yield of up to 30 μg of purified enzyme per milliliter of culture was obtained. The purified enzyme preparation showed properties identical to those previously reported in the literature.Keywords
This publication has 9 references indexed in Scilit:
- Beta‐lactamases and bacterial resistance to antibioticsMolecular Microbiology, 1995
- High specificity of cphA-encoded metallo-beta-lactamase from Aeromonas hydrophila AE036 for carbapenems and its contribution to beta-lactam resistanceAntimicrobial Agents and Chemotherapy, 1993
- An overview of the kinetic parameters of class B β-lactamasesBiochemical Journal, 1993
- The Aeromonas hydrophila cphA gene: molecular heterogeneity among class B metallo-beta-lactamasesJournal of Bacteriology, 1991
- Solubility as a Function of Protein Structure and Solvent ComponentsNature Biotechnology, 1990
- [6] Use of T7 RNA polymerase to direct expression of cloned genesPublished by Elsevier ,1990
- The structure of β-lactamasesPhilosophical Transactions of the Royal Society of London. B, Biological Sciences, 1980
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970