Journal
SCIENCE
Volume 346, Issue 6216, Pages 1525-1528Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1259680
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Funding
- NSF [CHE1306646]
- U. S. Department of Energy, Offices of Basic Energy Sciences and Biological and Environmental Research
- NIH
- Direct For Mathematical & Physical Scien
- Division Of Chemistry [1306646] Funding Source: National Science Foundation
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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-beta-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-beta-lactamase is functional in the Escherichia coli periplasm and enables the bacteria to survive treatment with ampicillin. In vivo screening of libraries has yielded a variant that displays a catalytic proficiency [(k(cat)/K-m)/k(uncat)] for ampicillin hydrolysis of 2.3 x 10(6) and features the emergence of a highly mobile loop near the active site, a key component of natural beta-lactamases to enable substrate interactions.
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