Several beta-lactamases, enzymes that play an important part in antibiotic resistance, have been purified by affinity chromatography on boronic acid gels. The procedure is rapid, appears to be selective for beta-lactamases, and allows a one-step purification of large amounts of enzyme from crude cell extracts. We have found the method useful for any beta-lactamase that is inhibited by boronic acids. Two kinds of boronic acid column have been prepared, the more hydrophobic one being reserved for those beta-lactamases that bind boronic acids relatively weakly. beta-Lactamase I from Bacillus cereus, P99 beta-lactamase and K 1 beta-lactamase from Gram-negative bacteria are among the better-known beta-lactamases that have been purified by this method. The procedure has also been used to purify a novel beta-lactamase from Pseudomonas maltophilia in high yield; the enzyme has an exceptionally broad substrate profile and hydrolyses monocyclic beta-lactams such as azthreonam and desthiobenzylpenicillin.
Research Article|July 15 1984
Purification of β-lactamases by affinity chromatography on phenylboronic acid-agarose
Biochem J (1984) 221 (2): 505-512.
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S J Cartwright, S G Waley; Purification of β-lactamases by affinity chromatography on phenylboronic acid-agarose. Biochem J 15 July 1984; 221 (2): 505–512. doi: https://doi.org/10.1042/bj2210505
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