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- Woon Ju Song
- Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0356, USA.
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- F. Akif Tezcan
- Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0356, USA.
書誌事項
- 公開日
- 2014-12-19
- DOI
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- 10.1126/science.1259680
- 公開者
- American Association for the Advancement of Science (AAAS)
この論文をさがす
説明
<jats:p> The generation of new enzymatic activities has mainly relied on repurposing the interiors of preexisting protein folds because of the challenge in designing functional, three-dimensional protein structures from first principles. Here we report an artificial metallo-β-lactamase, constructed via the self-assembly of a structurally and functionally unrelated, monomeric redox protein into a tetrameric assembly that possesses catalytic zinc sites in its interfaces. The designed metallo-β-lactamase is functional in the <jats:italic>Escherichia coli</jats:italic> periplasm and enables the bacteria to survive treatment with ampicillin. In vivo screening of libraries has yielded a variant that displays a catalytic proficiency [( <jats:italic>k</jats:italic> <jats:sub>cat</jats:sub> / <jats:italic>K</jats:italic> <jats:sub>m</jats:sub> )/ <jats:italic>k</jats:italic> <jats:sub>uncat</jats:sub> ] for ampicillin hydrolysis of 2.3 × 10 <jats:sup>6</jats:sup> and features the emergence of a highly mobile loop near the active site, a key component of natural β-lactamases to enable substrate interactions. </jats:p>
収録刊行物
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- Science
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Science 346 (6216), 1525-1528, 2014-12-19
American Association for the Advancement of Science (AAAS)
