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Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1973 Jan;3(1):68–73. doi: 10.1128/aac.3.1.68

Characterization of R Factor β-Lactamases by the Acidimetric Method

Fran A Rubin 1, David H Smith 1
PMCID: PMC444362  PMID: 4597709

Abstract

The properties and regulation of β-lactamases produced by certain R factors derived from ampicillin-resistant strains of Escherichia coli and Klebsiella were characterized. β-Lactamase activity was determined by the acidimetric method with phenol red as indicator. The sensitivity of this assay was increased by employing phosphate buffer at a final concentration of 0.4 mm; as little as 0.05 unit (1 unit equals the amount of β-lactamase that hydrolyzes 1 μmole of benzyl penicillin per hr at pH 7.6 and 25 C) could be detected. This assay is rapid and convenient and appears to be superior to other methods currently employed to assay R factor β-lactamases. Two classes of β-lactamases were distinguished on the basis of substrate profile, heat inactivation, and Km values. The regulation of most of these R factor β-lactamases, like certain other R factor enzymes, is subject to cyclic adenosine monophosphate-mediated catabolite repression.

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Selected References

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