Characterization of expression of a plant oleate desaturase in yeast

• Published in: Journal of the American Oil Chemists' Society 1998-01, Vol.75 (1), p.77-79
• Main Authors: Brown, A.P. (John Innes Centre, Norwich, UK.), Dann, R, Bowra, S, Hills, M.J
• Format: Article
• Language: English
• Subjects: Amino acids; Analytical, structural and metabolic biochemistry; Antibodies; Arabidopsis; ARABIDOPSIS THALIANA; Biological and medical sciences; Cell culture; Desaturase; Enzymes and enzyme inhibitors; Fatty acids; Fundamental and applied biological sciences. Psychology; Galactose; Genes; Lecithin; linoleate; Lipids; oleate; Oxidoreductases; Phosphatidylcholine; Phospholipids; SACCHAROMYCES CEREVISIAE; Yeast; Yeasts
• ISSN: 0003-021X; 1558-9331
• DOI: 10.1007/s11746-998-0014-y

We have expressed the oleate desaturase gene of Arabidopsis thaliana in Saccharomyces cerevisiae by means of a galactose‐inducible promoter. Transfer of yeast cells to galactose‐containing media and growth at 30°C resulted in the production of active oleate desaturase, as shown by the appearance of the diunsaturated octadecadienoic acid (18:2) and hexadecadienoic acid (16:2). The highest level of these two fatty acids was 46.2%. In induced cultures, 18:2 appeared before 16:2, and we show that both of these fatty acids were present first on phosphatidylcholine and then appeared in other phospholipids and a neutral lipid fraction. We raised antibodies against the C‐terminal 100 amino acids of oleate desaturase and demonstrated that this protein was in the microsomal fraction of induced cells.