Purification of a Jojoba Embryo Wax Synthase, Cloning of Its cDNA, and Production of High Levels of Wax in Seeds of Transgenic Arabidopsis

• Published in: Plant physiology (Bethesda) 2000-03, Vol.122 (3), p.645-655
• Main Authors: Kathryn D. Lardizabal, Metz, James G., Tetsuo Sakamoto, Hutton, William C., Pollard, Michael R., Lassner, Michael W.
• Format: Article
• Language: English
• Subjects: Acyltransferases - genetics; Acyltransferases - isolation & purification; Agronomy. Soil science and plant productions; Alcohols; Amino Acid Sequence; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis thaliana; b-ketoacyl-CoA synthase; Base Sequence; Biochemistry and Macromolecular Structure; Biological and medical sciences; Cloning, Molecular; Complementary DNA; DNA Primers - genetics; DNA, Complementary - genetics; DNA, Plant - genetics; Economic plant physiology; Embryos; Enzymes; Esters; Fatty acids; Fatty alcohols; Fundamental and applied biological sciences. Psychology; Gene Expression; Lipids; Metabolism; Molecular Sequence Data; Nutrition. Photosynthesis. Respiration. Metabolism; Open reading frames; Plant Oils - chemistry; Plant Oils - metabolism; Plant physiology and development; Plants - embryology; Plants - enzymology; Plants - genetics; Plants, Genetically Modified; Seeds; Seeds - genetics; Seeds - metabolism; Sequence Homology, Amino Acid; Simmondsia chinensis; Waxes; Waxes - chemistry; Waxes - metabolism
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.122.3.645

Wax synthase (WS, fatty acyl-coenzyme A [coA]: fatty alcohol acyltransferase) catalyzes the final step in the synthesis of linear esters (waxes) that accumulate in seeds of jojoba (Simmondsia chinensis). We have characterized and partially purified this enzyme from developing jojoba embryos. A protein whose presence correlated with WS activity during chromatographic fractionation was identified and a cDNA encoding that protein was cloned. Seed-specific expression of the cDNA in transgenic Arabidopsis conferred high levels of WS activity on developing embryos from those plants. The WS sequence has significant homology with several Arabidopsis open reading frames of unknown function. Wax production in jojoba requires, in addition to WS, a fatty acyl-CoA reductase (FAR) and an efficient fatty acid elongase system that forms the substrates preferred by the FAR. We have expressed the jojoba WS cDNA in Arabidopsis in combination with cDNAs encoding the jojoba FAR and a β-ketoacyl-CoA synthase (a component of fatty acid elongase) from Lunaria annua. 13C-Nuclear magnetic resonance analysis of pooled whole seeds from transgenic plants indicated that as many as 49% of the oil molecules in the seeds were waxes. Gas chromatography analysis of transmethylated oil from individual seeds suggested that wax levels may represent up to 70% (by weight) of the oil present in those seeds.