+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Characterization and engineering of the bifunctional N- and O-glucosyltransferase involved in xenobiotic metabolism in plants



Characterization and engineering of the bifunctional N- and O-glucosyltransferase involved in xenobiotic metabolism in plants



Proceedings of the National Academy of Sciences of the United States of America 104(51): 20238-20243



The glucosylation of pollutant and pesticide metabolites in plants controls their bioactivity and the formation of subsequent chemical residues. The model plant Arabidopsis thaliana contains >100 glycosyltransferases (GTs) dedicated to small-molecule conjugation and, whereas 44 of these enzymes catalyze the O-glucosylation of chlorinated phenols, only one, UGT72B1, shows appreciable N-glucosylating activity toward chloroanilines. UGT72B1 is a bifunctional O-glucosyltransferase (OGT) and N-glucosyltransferase (NGT). To investigate this unique dual activity, the structure of the protein was solved, at resolutions up to 1.45 A, in various forms including the Michaelis complex with intact donor analog and trichlorophenol acceptor. The catalytic mechanism and basis for O/N specificity was probed by mutagenesis and domain shuffling with an orthologous enzyme from Brassica napus (BnUGT), which possesses only OGT activity. Mutation of BnUGT at just two positions (D312N and F315Y) installed high levels of NGT activity. Molecular modeling revealed the connectivity of these residues to H19 on UGT72B1, with its mutagenesis exclusively defining NGT activity in the Arabidopsis enzyme. These results shed light on the conjugation of nonnatural substrates by plant GTs, highlighting the catalytic plasticity of this enzyme class and the ability to engineer unusual and desirable transfer to nitrogen-based acceptors.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 018540566

Download citation: RISBibTeXText

PMID: 18077347

DOI: 10.1073/pnas.0706421104


Related references

Conjugating enzymes involved in xenobiotic metabolism of organic xenobiotics in plants. International Journal of Phytoremediation 4(4): 247-265, 2002

Multiple xenobiotic-inducible P450s are involved in alkoxycoumarin and alkoxyresorufin metabolism in higher plants. Plant Cell & Environment 18(5): 523-533, 1995

Transgenic rice plants expressing human p450 genes involved in xenobiotic metabolism for phytoremediation. Journal of Molecular Microbiology and Biotechnology 15(2-3): 212-219, 2008

Design and characterization of a novel "family-shuffling" technology adapted to membrane enzyme: application to P450s involved in xenobiotic metabolism. Advances in Experimental Medicine and Biology 500: 319-322, 2001

Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus. Bmc Microbiology 16(1): 280, 2016

Preparation and characterization of subcellular fractions from the intestinal mucosa of the northern pike esox lucius with special emphasis on enzymes involved in xenobiotic metabolism. Biochimica et Biophysica Acta 838(2): 277-289, 1985

Characterization of a bifunctional O- and N-glucosyltransferase from Vitis vinifera in glucosylating phenolic compounds and 3,4-dichloroaniline in Pichia pastoris and Arabidopsis thaliana. Plos one 8(11): E80449, 2013

Identification of glucosyltransferase genes involved in sinapate metabolism and lignin synthesis in Arabidopsis. Journal of Biological Chemistry 276(6): 4344-4349, 2001

Engineering isoflavone metabolism with an artificial bifunctional enzyme. Planta 224(3): 496-507, 2006

Cloning and characterization of a glucosyltransferase from Crocus sativus stigmas involved in flavonoid glucosylation. Bmc Plant Biology 9: 109, 2009

Isolation and characterisation of carboxylesterases involved in xenobiotic metabolism in rice. The BCPC Conference: Weeds, 2001, Volume 1 and Volume 2 Proceedings of an international conference held at the Brighton Hilton Metropole Hotel, Brighton, UK, 12-15 November 2001 : 337-340, 2001

Human and rat liver microsomal cytochrome p 450 involved in xenobiotic metabolism. Research and Practice in Forensic Medicine 28: 79-84, 1985

Characterization of a glucosyltransferase enzyme involved in the formation of kaempferol and quercetin sophorosides in Crocus sativus. Plant Physiology 159(4): 1335-1354, 2012

Purification and characterization of glucosyltransferase and glucanotransferase involved in the production of cyclic tetrasaccharide in Bacillus globisporus C11. Bioscience Biotechnology and Biochemistry 66(9): 1806-1818, 2002

Xenobiotic metabolism in plants. Drug Metabolism from Microbe to Man: 91-217, 1977