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PMID: 6332621 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification of beta-lactamases by affinity chromatography on phenylboronic acid-agarose.

The Biochemical journal ·Vol. 221 ·No. 2 ·1984-07-15 ·Pages 505-12

Cartwright SJ, Waley SG

Abstract

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.

MeSH Terms
Chromatography, Affinity/methods Isoelectric Focusing Pseudomonas/enzymology Sepharose/analogs & derivatives Substrate Specificity beta-Lactamases/isolation & purification
Chemicals
phenylboronic acid-sepharose Sepharose beta-Lactamases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cartwright S J
Waley S G
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31 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2210505
Published
1984-07-15
Pages
505-12
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1144066
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2210505 PMC Full Text View journal details
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