Isolation of a glucosyltransferase from Arabidopsis thaliana active in the metabolism of the persistent pollutant 3,4-dichloroaniline.
The pollutant 3,4-dichloroaniline (DCA) was rapidly detoxified by glucosylation in Arabidopsis thaliana root cultures, with the N-beta-d-glucopyranosyl-DCA exported into the medium. The N-glucosyltransferase (N-GT) responsible for this activity was purified from Arabidopsis suspension cultures and the resulting 50 kDa polypeptide analysed by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) following tryptic digestion. The protein was identified as GT72B1. The GT was cloned and the purified recombinant enzyme shown to be highly active in conjugating DCA and 2,4,5-trichlorophenol, as well as several other chlorinated phenols and anilines, demonstrating both N-GT and O-GT activity. GT72B1 showed little activity towards natural products with the exception of the tyrosine catabolite 4-hydroxyphenylpyruvic acid. Both O-GT and N-GT activities were enhanced in both plants and cultures treated with herbicide safeners, demonstrating the chemical inducibility of this detoxification system in Arabidopsis.
Loutre
Caroline
C
Crop Protection Group, School of Biological and Biomedical Sciences, University of Durham, UK.
Dixon
David P
DP
Brazier
Melissa
M
Slater
Malcolm
M
Cole
David J
DJ
Edwards
Robert
R
D000595:Amino Acid Sequence; D000814:Aniline Compounds / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004785:Environmental Pollutants / Q000378:metabolism*; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D066298:In Vitro Techniques; D008658:Inactivation, Metabolic; D008969:Molecular Sequence Data; D018517:Plant Roots / Q000378:metabolism; D013379:Substrate Specificity
D000814:Aniline Compounds; D004785:Environmental Pollutants; C014464:3,4-dichloroaniline; D016695:Glycosyltransferases