Discovery and Directed Evolution of a Glyphosate Tolerance Gene

作者：Linda A. Castle, Daniel L. Siehl, Rebecca Gorton, Phillip A. Patten, Yong Hong Chen, Sean Bertain, Hyeon‐Je Cho, Nicholas B. Duck, James F. H. Wong, Donglong Liu, Michael Lassner · 发表于：Science · 年份：2004 · DOI：10.1126/science.1096770 · 被引用次数：328 · 研究领域：CRISPR and Genetic Engineering、Plant tissue culture and regeneration、Plant nutrient uptake and metabolism

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.