Influence of sulfobetaines on the stability of the Citrobacterdiversus ULA-27 beta-lactamase.
The activity and stability of beta-lactamase from Citrobacter diversus ULA-27 have been investigated in the presence of different ionic and zwitterionic surfactants. All the sulfobetaine surfactants tested allow the enzyme to retain its full activity, but the best stabilizing effect is greatly dependent on their structure. Very little variations on the monomer headgroup can significantly reduce enzyme deactivation or speed up the loss of activity with respect to buffer alone. The whole hydrophobic/hydrophilic balance on the headgroup seems to have a determining role in preserving beta-lactamase activity and structure. The presence of zwitterionic surfactants stabilizes the protein conformation toward denaturation by urea and low-temperature inactivation. Similar experiments were performed in the presence of other two zwitterionic surfactants, an amine oxide, dimethylmyristylamine oxide (DMMAO) and a carboxybetaine, cetyldimethylammonium methanecarboxylate (CB1-16). The former stabilizes the enzyme even better than the sulfobetaines, the latter quickly deactivates it. Therefore, the factors responsible for beta-lactamase stabilization are dependent not only on the zwitterionic nature of the surfactant headgroup but also specific interactions between the surfactant and the protein may be important.
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Dipartimento di Chimica, Ingegneria Chimica e Materiali, Università dell'Aquila, Via Vetoio, Coppito 2, 67100 L'Aquila, Italia.
Reale
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Amicosante
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Profio
P D
PD
Germani
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Savelli
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G
D001622:Betaine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D002954:Citrobacter / Q000201:enzymology* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004795:Enzyme Stability / Q000187:drug effects; D006358:Hot Temperature; D007700:Kinetics; D008823:Micelles; D011489:Protein Denaturation; D012492:Salts; D012996:Solutions; D013501:Surface-Active Agents / Q000494:pharmacology*; D014508:Urea / Q000737:chemistry; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000737:chemistry / Q000302:isolation & purification
D004791:Enzyme Inhibitors; D008823:Micelles; D012492:Salts; D012996:Solutions; D013501:Surface-Active Agents; D065093:beta-Lactamase Inhibitors; D001622:Betaine; D014508:Urea; D001618:beta-Lactamases