Assays for beta-lactamase activity and inhibition

• Published in: Methods in molecular medicine Vol. 142; p. 239
• Main Authors: Viswanatha, Thammaiah, Marrone, Laura, Goodfellow, Valerie, Dmitrienko, Gary I
• Format: Journal Article
• Language: English
• Published: United States 2008
• Subjects: Bacterial Proteins - analysis; Bacterial Proteins - antagonists & inhibitors; Bacterial Proteins - isolation & purification; beta-Lactamase Inhibitors; beta-Lactamases - analysis; beta-Lactamases - isolation & purification; beta-Lactams - pharmacology; Chelating Agents - analysis; Enzyme Inhibitors - analysis; Enzyme Inhibitors - pharmacology; Enzyme Stability; Escherichia coli - enzymology; Humans; Kinetics; Spectrophotometry
• ISSN: 1543-1894
• DOI: 10.1007/978-1-59745-246-5_19

The ability, either innate or acquired, to produce beta-lactamases, enzymes capable of hydrolyzing the endocyclic peptide bond in beta-lactam antibiotics, would appear to be a primary contributor to the ever-increasing incidences of resistance to this class of antibiotics. To date, four distinct classes, A, B, C, and D, of beta-lactamases have been identified. Of these, enzymes in classes A, C, and D utilize a serine residue as a nucleophile in their catalytic mechanism while class B members are Zn2+-dependent for their function. Efforts have been and still continue to be made toward the development of potent inhibitors of these enzymes as a means to ensure the efficacy of beta-lactam antibiotics in clinical medicine. This chapter concerns procedures for the evaluation of the catalytic activity of beta-lactamases as a means to screen compounds for their inhibitory potency.