Plant Physiology Preview
Published on August 7, 2003; 10.1104/pp.102.019240
Received December 17, 2002
Returned for revision January 29, 2003
Accepted May 12, 2003
CYP83A1 and CYP83B1, Two Nonredundant Cytochrome P450 Enzymes Metabolizing Oximes in the Biosynthesis of Glucosinolates in Arabidopsis
Peter Naur , Bent Larsen Petersen , Michael Dalgaard Mikkelsen , Søren Bak , Hasse Rasmussen , Carl Erik Olsen , and Barbara Ann Halkier *
Plant Biochemistry Laboratory (P.N., B.L.P., M.D.M., S.B., B.A.H.) and Department of Chemistry (C.E.O.), Center for Molecular Plant Physiology, The Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark; and IACR-Rothamsted, Harpenden AL5 2JQ, United Kingdom (H.R.)
* Corresponding author; email: bah{at}kvl.dk.
In the glucosinolate pathway, the postoxime enzymes have been proposed to have low specificity for the side chain and high specificity for the functional group. Here, we provide biochemical evidence for the functional role of the two cytochromes P450, CYP83A1 and CYP83B1, from Arabidopsis in oxime metabolism in the biosynthesis of glucosinolates. In a detailed analysis of the substrate specificities of the recombinant enzymes heterologously expressed in yeast (Saccharomyces cerevisiae), we show that aliphatic oximes derived from chain-elongated homologs of methionine are efficiently metabolized by CYP83A1, whereas CYP83B1 metabolizes these substrates with very low efficiency. Aromatic oximes derived from phenylalanine, tryptophan, and tyrosine are metabolized by both enzymes, although CYP83B1 has higher affinity for these substrates than CYP83A1, particularly in the case of indole-3-acetaldoxime, where there is a 50-fold difference in Km value. The data show that CYP83A1 and CYP83B1 are nonredundant enzymes under physiologically normal conditions in the plant. The ability of CYP83A1 to metabolize aromatic oximes, albeit at small levels, explains the presence of indole glucosinolates at various levels in different developmental stages of the CYP83B1 knockout mutant, rnt1-1. Plants overexpressing CYP83B1 contain elevated levels of aliphatic glucosinolates derived from methionine homologs, whereas the level of indole glucosinolates is almost constant in the overexpressing lines. Together with the previous characterization of the members of the CYP79 family involved in oxime production, this work provides a framework for metabolic engineering of glucosinolates and for further dissection of the glucosinolate pathway.
This article has been cited by other articles:
|
|
|
|
 
E. G. Bakker, M. B. Traw, C. Toomajian, M. Kreitman, and J. Bergelson
Low Levels of Polymorphism in Genes That Control the Activation of Defense Response in Arabidopsis thaliana
Genetics,
April 1, 2008;
178(4):
2031 - 2043.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Nafisi, S. Goregaoker, C. J. Botanga, E. Glawischnig, C. E. Olsen, B. A. Halkier, and J. Glazebrook
Arabidopsis Cytochrome P450 Monooxygenase 71A13 Catalyzes the Conversion of Indole-3-Acetaldoxime in Camalexin Synthesis
PLANT CELL,
June 1, 2007;
19(6):
2039 - 2052.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Textor, J.-W. de Kraker, B. Hause, J. Gershenzon, and J. G. Tokuhisa
MAM3 Catalyzes the Formation of All Aliphatic Glucosinolate Chain Lengths in Arabidopsis
Plant Physiology,
May 1, 2007;
144(1):
60 - 71.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Duan, M.-Y. Huang, K. Palacio, and M. A. Schuler
Variations in CYP74B2 (Hydroperoxide Lyase) Gene Expression Differentially Affect Hexenal Signaling in the Columbia and Landsberg erecta Ecotypes of Arabidopsis
Plant Physiology,
November 1, 2005;
139(3):
1529 - 1544.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Zhao, J. C. Craig, H. E. Petzold, A. W. Dickerman, and E. P. Beers
The Xylem and Phloem Transcriptomes from Secondary Tissues of the Arabidopsis Root-Hypocotyl
Plant Physiology,
June 1, 2005;
138(2):
803 - 818.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. M. Fraser, L. W. Rider, and C. Chapple
An Expression and Bioinformatics Analysis of the Arabidopsis Serine Carboxypeptidase-Like Gene Family
Plant Physiology,
June 1, 2005;
138(2):
1136 - 1148.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Kristensen, M. Morant, C. E. Olsen, C. T. Ekstrom, D. W. Galbraith, B. Lindberg Moller, and S. Bak
Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome
PNAS,
February 1, 2005;
102(5):
1779 - 1784.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. L. Celenza, J. A. Quiel, G. A. Smolen, H. Merrikh, A. R. Silvestro, J. Normanly, and J. Bender
The Arabidopsis ATR1 Myb Transcription Factor Controls Indolic Glucosinolate Homeostasis
Plant Physiology,
January 1, 2005;
137(1):
253 - 262.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. R. Nelson, M. A. Schuler, S. M. Paquette, D. Werck-Reichhart, and S. Bak
Comparative Genomics of Rice and Arabidopsis. Analysis of 727 Cytochrome P450 Genes and Pseudogenes from a Monocot and a Dicot
Plant Physiology,
June 1, 2004;
135(2):
756 - 772.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
