Abstract
Metallo-β-lactamase from Bacillus anthracis (Bla2) catalyzes the hydrolysis of β-lactam antibiotics which are commonly prescribed to combat bacterial infections. Bla2 contributes to the antibiotic resistance of this bacterium. An understanding of it is necessary to design potential inhibitors that can be introduced with current antibiotics for effective eradication of anthrax infections. We have purified Bla2 using Ni2+-affinity chromatography with over 140-fold increase in activity with a yield of 3.5%. The final specific activity was 19,000 units/mg. Purified Bla2 displays different K m , V max , and (k cat /K M) with penicillin G and cephalexin as substrates and is also sensitive to pH, with maximum activity between pH 7.0–9.0. The IC50 (50% inhibition concentration) value of EDTA against Bla2 is 630 nM, which can be understood by observing its three-dimensional interaction with the enzyme.
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Acknowledgments
This work was in part funded by grants from the Young Investigator Development Program and the University Research Committee at Baylor University. The authors are grateful to Dr. Timothy Palzkill (Baylor College of Medicine) for supplying the plasmid encoding the metallo-β-lactamase from B. anthracis.
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Sara R. Schlesinger and Sang Gon Kim contributed equally to this work.
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Schlesinger, S.R., Kim, S.G., Lee, JS. et al. Purification development and characterization of the zinc-dependent metallo-β-lactamase from Bacillus anthracis . Biotechnol Lett 33, 1417–1422 (2011). https://doi.org/10.1007/s10529-011-0569-9
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DOI: https://doi.org/10.1007/s10529-011-0569-9