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Applied and Environmental Microbiology, August 2005, p. 4771-4776, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4771-4776.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel AroA with High Tolerance to Glyphosate, Encoded by a Gene of Pseudomonas putida 4G-1 Isolated from an Extremely Polluted Environment in China

Yi-Cheng Sun,1 Yan-Cheng Chen,1 Zhe-Xian Tian,1 Feng-Mei Li,1 Xin-Yue Wang,1 Jing Zhang,1 Zhen-Long Xiao,1 Min Lin,2 Niamh Gilmartin,3 David N. Dowling,3 and Yi-Ping Wang1*

National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University,1 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China,2 Department of Science and Health, Institute of Technology Carlow, Kilkenny Road, Carlow, Ireland3

Received 1 September 2004/ Accepted 24 February 2005

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 Km[PEP], Km[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.

* Corresponding author. Mailing address: National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100081, People's Republic of China. Phone: (86) 10-6275 8490. Fax: (86) 10-6275 6325. E-mail: wangyp{at}pku.edu.cn.

Applied and Environmental Microbiology, August 2005, p. 4771-4776, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4771-4776.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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