Discovery and directed evolution of a glyphosate tolerance gene

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 transg...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2004-05, Vol.304 (5674), p.1151-1154
Main Authors: Castle, L.A, Siehl, D.L, Gorton, R, Patten, P.A, Chen, Y.H, Bertain, S, Cho, H.J, Duck, N, Wong, J, Liu, D
Format: Article
Language:English
Subjects:
Acetylation
Acetyltransferases - chemistry
Acetyltransferases - genetics
Acetyltransferases - metabolism
acyltransferases
Amino Acid Sequence
Amino acids
Arabidopsis thaliana
Bacillus - enzymology
Bacillus licheniformis
Botany
Catalysis
Corn
Crops
Directed Molecular Evolution
DNA Shuffling
Drug Resistance
enzyme activity
Enzymes
Escherichia coli - genetics
gene expression
Gene Library
gene shuffling
gene transfer
Genetic screening
Genetic Variation
Glycine - analogs & derivatives
Glycine - metabolism
Glycine - toxicity
Glyphosate
glyphosate N-acetyltransferase
Herbicide resistance
Herbicides
Herbicides - metabolism
Herbicides - toxicity
Kinetics
Libraries
Molecular Sequence Data
multigene shuffling
Mutagenesis
Nicotiana - drug effects
Nicotiana - genetics
Nicotiana - growth & development
Nicotiana tabacum
Plant growth
Plants
Plants, Genetically Modified - drug effects
Plants, Genetically Modified - genetics
protein engineering
recombinant proteins
Recombinant Proteins - metabolism
Recombination, Genetic
structural genes
tobacco
Transformation, Genetic
Transgenic plants
Zea mays
Zea mays - drug effects
Zea mays - genetics
Zea mays - growth & development
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Summary: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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.1096770