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J. Biol. Chem., Vol. 280, Issue 45, 37742-37746, November 11, 2005

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An Active Enzyme Constructed from a 9-Amino Acid Alphabet^*

From the Laboratory of Organic Chemistry, Swiss Federal Institute of Technology, ETH Hönggerberg, CH-8093 Zürich, Switzerland

Nature employs a set of 20 amino acids to produce a repertoire of protein structures endowed with sophisticated functions. Here, we combined design and selection to create an enzyme composed entirely from a set of only 9 amino acids that can rescue auxotrophic cells lacking chorismate mutase. The simplified protein captures key structural features of its natural counterpart but appears to be somewhat less stable and more flexible. The potential of a dramatically reduced amino acid alphabet to produce an active catalyst supports the notion that primordial enzymes may have possessed low amino acid diversity and suggests that combinatorial engineering strategies, such as the one used here, may be generally applied to create enzymes with novel structures and functions.

Received for publication, July 5, 2005 , and in revised form, August 15, 2005.

^* This work was supported by the Swiss Federal Institute of Technology and the Schweizerischer Nationalfonds. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains a supplemental table and three supplemental figures.

¹ To whom correspondence should be addressed: Swiss Federal Institute of Technology, ETH Hönggerberg, CH-8093 Zürich, Switzerland. Tel.: 41-1-632-3176; Fax: 41-1-632-1486; E-mail: hilvert{at}org.chem.ethz.ch.

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