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Science 21 May 2004:
Vol. 304. no. 5674, pp. 1151 - 1154
DOI: 10.1126/science.1096770
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Reports

Discovery and Directed Evolution of a Glyphosate Tolerance Gene

Linda A. Castle,1* Daniel L. Siehl,1 Rebecca Gorton,1 Phillip A. Patten,2 Yong Hong Chen,2 Sean Bertain,1 Hyeon-Je Cho,1 Nicholas Duck,3{dagger} James Wong,3 Donglong Liu,3 Michael W. Lassner1

The herbicide glyphosate is effectively detoxified by N-acetylation. We screened a collection of microbial isolates and discovered enzymes exhibiting glyphosate N-acetyltransferase (GAT) activity. Kinetic properties of the discovered enzymes were insufficient to confer glyphosate tolerance to transgenic organisms. Eleven iterations of DNA shuffling improved enzyme efficiency by nearly four orders of magnitude from 0.87 mM–1 min–1 to 8320 mM–1 min–1. From the fifth iteration and beyond, GAT enzymes conferred increasing glyphosate tolerance to Escherichia coli, Arabidopsis, tobacco, and maize. Glyphosate acetylation provides an alternative strategy for supporting glyphosate use on crops.

1 Verdia, Inc. Redwood City, CA 94063, USA.
2 Maxygen Inc., Redwood City, CA 94063, USA.
3 Pioneer Hi-Bred International, Inc., Johnston, IA 50131, USA.



{dagger} Present address: Athenix Corporation, Research Triangle Park, NC 27709, USA.

* To whom correspondence should be addressed. E-mail: linda.castle{at}verdiainc.com.

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