The activity of the dinuclear cobalt-β-lactamase from Bacillus cereus in catalysing the hydrolysis of β-lactams

Adriana Badarau, Christian Damblon, Michael I. Page

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Metallo-β-lactamases are native zinc enzymes that catalyse the hydrolysis of β-lactam antibiotics, but are also able to function with cobalt(II) and require one or two metal-ions for catalytic activity. The hydrolysis of cefoxitin, cephaloridine and benzylpenicillin catalysed by CoBcII (cobalt-substituted β-lactamase from Bacillus cereus) has been studied at different pHs and metal-ion concentrations. An enzyme group of pKa 6.52 ± 0.1 is found to be required in its deprotonated form for metal-ion binding and catalysis. The species that results from the loss of one cobalt ion from the enzyme has no significant catalytic activity and is thought to be the mononuclear CoBcII. It appears that dinuclear CoBcII is the active form of the enzyme necessary for turnover, while the mononuclear CoBcII is only involved in substrate binding. The cobalt-substituted enzyme is a more efficient catalyst than the native enzyme for the hydrolysis of some β-lactam antibiotics suggesting that the role of the metal-ion is predominantly to provide the nucleophilic hydroxide, rather than to act as a Lewis acid to polarize the carbonyl group and stabilize the oxyanion tetrahedral intermediate.

LanguageEnglish
Pages197-203
Number of pages7
JournalBiochemical Journal
Volume401
Issue number1
Early online date11 Sep 2006
DOIs
Publication statusPublished - 1 Jan 2007

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Bacillus cereus
beta-Lactams
beta-Lactamases
Cobalt
Hydrolysis
Metal ions
Ions
Enzymes
Metals
Catalyst activity
Cephaloridine
Anti-Bacterial Agents
Cefoxitin
Lewis Acids
Penicillin G
Catalysis
Zinc
Catalysts
Substrates

Cite this

Badarau, Adriana ; Damblon, Christian ; Page, Michael I. / The activity of the dinuclear cobalt-β-lactamase from Bacillus cereus in catalysing the hydrolysis of β-lactams. In: Biochemical Journal. 2007 ; Vol. 401, No. 1. pp. 197-203.
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The activity of the dinuclear cobalt-β-lactamase from Bacillus cereus in catalysing the hydrolysis of β-lactams. / Badarau, Adriana; Damblon, Christian; Page, Michael I.

In: Biochemical Journal, Vol. 401, No. 1, 01.01.2007, p. 197-203.

Research output: Contribution to journalArticle

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