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Characterization of expression of a plant oleate desaturase in yeast
Author(s) -
Brown Adrian P.,
Dann Rachel,
Bowra Steve,
Hills Matthew J.
Publication year - 1998
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1007/s11746-998-0014-y
Subject(s) - yeast , biochemistry , saccharomyces cerevisiae , phosphatidylcholine , fatty acid desaturase , microsome , galactose , arabidopsis thaliana , fatty acid , stearoyl coa desaturase , amino acid , chemistry , biology , gene expression , gene , polyunsaturated fatty acid , enzyme , mutant , phospholipid , membrane
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.