Induction of beta-lactamase from Shigella flexneri UCSF-129: purification by affinity chromatography and some properties.
Lincomycin, an inhibitor of protein synthesis, was an excellent inductor of beta-lactamase, and its total activity and specific activity were increased 2.5- and 3.6-fold respectively. The beta-lactamase produced by Shigella flexneri UCSF-129 was purified by affinity chromatography on phenylboronic acid-agarose with a type B column using an hydrophobic spacer arm (6-aminohexanoic acid activated with succinimide). The yield and specific activity were 96% and 29,283 U/mg, respectively. These were 1.7- and 3.8-fold higher, respectively, than those obtained by the traditional method using ion-exchange chromatography and gel filtration. The working-time was reduced to a third, and the enzyme preparation was shown to be pure by several criteria. From nine divalent cations assayed, only Sn(II) inhibited the enzyme by 74%, and the chloride ion did not have any effect on enzyme activity.
Campos
M
M
Department of Chemistry, Faculty of Science, University of Concepción, Chile.
González
H
H
Alarcón
M A
MA
Sánchez
R
R
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002846:Chromatography, Affinity; D004790:Enzyme Induction / Q000187:drug effects; D008034:Lincomycin / Q000494:pharmacology; D012763:Shigella flexneri / Q000187:drug effects / Q000201:enzymology*; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism
D000900:Anti-Bacterial Agents; D008034:Lincomycin; D001618:beta-Lactamases