Download | - View author's version: A FAD2 homologue from Lesquerella lindheimeri has predominantly fatty acid hydroxylase activity (PDF, 5.8 MiB)
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DOI | Resolve DOI: https://doi.org/10.1016/j.plantsci.2007.03.015 |
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Author | Search for: Dauk, Melanie; Search for: Lam, Patricia1; Search for: Kunst, Ljerk1; Search for: Smith, Mark A. |
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Affiliation | - National Research Council of Canada. NRC Plant Biotechnology Institute
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Format | Text, Article |
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Subject | lesquerella lindheimeri; fatty acid hydroxylase; ricinoleic acid; hydroxy fatty acid; seed oil content; single seed TLC |
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Abstract | A genomic sequence encoding a polypeptide with 91% sequence identity to the Lesquerella fendleri bifunctional oleate 12-hydroxylase:desaturase was amplified by PCR from Lesquerella lindheimeri. Expression of the gene in the yeast Saccharomyces cerevisiae resulted in the synthesis of ricinoleic acid and very low levels of di-unsaturated fatty acids. Comparison of the amino acid sequences of the L. lindheimeri and castor bean oleate 12-hydroxylase enzymes to those of the L. fendleri bifunctional oleate 12-hydroxylase:desaturase and oleate 12-desaturase enzymes from 32 diverse species identified a single amino acid (M295) that was conserved in the hydroxylases and different but also conserved in the desaturases and the bifunctional enzyme. Site-directed mutagenesis indicated that this residue was most likely not involved in determining the catalytic outcome of the hydroxylation/desaturation reaction. Transformation of an Arabidopsis fad2/fae1 mutant line with the L. lindheimeri hydroxylase gave further evidence that this enzyme, like the castor oleate 12-hydroxylase, is primarily a fatty acid hydroxylase and should not be considered bifunctional. Total hydroxy fatty acid content of up to 18% of seed fatty acids was measured in homozygous transformants. Lines with the highest hydroxy fatty acid content showed significant reduction in total oil content. |
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Publication date | 2007-04-04 |
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Publisher | Elsevier |
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In | |
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Language | English |
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Peer reviewed | Yes |
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NPARC number | 23002566 |
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Export citation | Export as RIS |
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Report a correction | Report a correction (opens in a new tab) |
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Record identifier | 6c69af17-0d37-475c-8c4f-2d34d1cde352 |
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Record created | 2017-11-30 |
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Record modified | 2020-05-10 |
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