title: Novel AroA with high tolerance to glyphosate, encoded by a gene of Pseudomonas putida 4G-1 isolated from an extremely polluted environment in China.
abstract: Glyphosate has been used globally as a safe herbicide for weed control. It inhibits 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase (AroA), which is a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants. A Pseudomonas putida strain, 4G-1, was isolated from a soil heavily contaminated by glyphosate in China. Its AroA-encoding gene (aroA) has been cloned, sequenced, and expressed in Escherichia coli. Phylogenetic analysis revealed that this AroA belongs neither to class I nor to class II AroA enzymes. When compared with E. coli AroA, 4G-1 AroA shows similar values for K(m)[PEP], K(m)[S3P], and specific enzyme activity. Moreover, 4G-1 AroA exhibits high tolerance to glyphosate, which indicates a protein with a high potential for structural and functional studies of AroA in general and its potential usage for the generation of transgenic crops resistant to the herbicide.
authors.affiliation: National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100081, People's Republic of China.
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Sun
authors.forename: Yan-Cheng
authors.initials: YC
authors.lastname: Chen
authors.forename: Zhe-Xian
authors.initials: ZX
authors.lastname: Tian
authors.forename: Feng-Mei
authors.initials: FM
authors.lastname: Li
authors.forename: Xin-Yue
authors.initials: XY
authors.lastname: Wang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhen-Long
authors.initials: ZL
authors.lastname: Xiao
authors.forename: Min
authors.initials: M
authors.lastname: Lin
authors.forename: Niamh
authors.initials: N
authors.lastname: Gilmartin
authors.forename: David N
authors.initials: DN
authors.lastname: Dowling
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Wang
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002681:China; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006540:Herbicides / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016958:Pseudomonas putida / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D012989:Soil Pollutants / Q000494:pharmacology; D000097797:Glyphosate
chemical_list: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D005998:Glycine; D006540:Herbicides; D012989:Soil Pollutants; D000097797:Glyphosate