An alternative to the insensitive target approach used to develop glyphosate tolerant

crops is to engineer crops to produce an enzyme capable of detoxifying the herbicide.

Phosphinothricin acetyltransferase (PAT or BAR) from Streptomyces detoxifies
phosphinothricin- or bialaphos-based herbicides by adding an acetyl group
2
. This trait
is approved for use in the United States in canola, chicory, maize, rice,
cotton,soybean, and sugarbeet. Because detoxification by acetylation is
straightforward, requires no cofactors, and has proven to be a successful strategy for
phosphinothricin detoxification, we sought an enzyme capable of carrying out the N-
acetylation of glyphosate. Previous research demonstrated that N-acetylglyphosate is
not herbicidal.
By utilizing the sensitivity and specificity of mass spectrometry (MS) to query natural
abilities of common microbes, we were able to discover glyphosate acetyltransferase
(GAT) enzymes capable of detoxifying glyphosate by N-acetylation. Because native
enzymes exhibited poor kinetic properties, we used DNA shuffling
3-5
to create
improved enzymes that exhibit higher turnover rates and increased specificity for
glyphosate. The improved N-acetyl- transferase genes (gat), when expressed in plants,
lead to a robust glyphosate tolerant phenotype as described in detail recently
6
(Fig. 1).