Purification of a jojoba embryo fatty acyl-coenzyme A reductase and expression of its cDNA in high erucic acid rapeseed.
The jojoba (Simmondsia chinensis) plant produces esters of long-chain alcohols and fatty acids (waxes) as a seed lipid energy reserve. This is in contrast to the triglycerides found in seeds of other plants. We purified an alcohol-forming fatty acyl-coenzyme A reductase (FAR) from developing embryos and cloned the cDNA encoding the enzyme. Expression of a cDNA in Escherichia coli confers FAR activity upon those cells and results in the accumulation of fatty alcohols. The FAR sequence shows significant homology to an Arabidopsis protein of unknown function that is essential for pollen development. When the jojoba FAR cDNA is expressed in embryos of Brassica napus, long-chain alcohols can be detected in transmethylated seed oils. Resynthesis of the gene to reduce its A plus T content resulted in increased levels of alcohol production. In addition to free alcohols, novel wax esters were detected in the transgenic seed oils. In vitro assays revealed that B. napus embryos have an endogenous fatty acyl-coenzyme A: fatty alcohol acyl-transferase activity that could account for this wax synthesis. Thus, introduction of a single cDNA into B. napus results in a redirection of a portion of seed oil synthesis from triglycerides to waxes.
Metz
J G
JG
Calgene Campus, Monsanto, 1920 Fifth Street, Davis, California 95616, USA. jim.metz@monsanto.com
Pollard
M R
MR
Anderson
L
L
Hayes
T R
TR
Lassner
M W
MW
D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004883:Erucic Acids / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D010938:Plant Oils / Q000378:metabolism; D010944:Plants / Q000196:embryology / Q000201:enzymology* / Q000235:genetics*; D030821:Plants, Genetically Modified
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D004883:Erucic Acids; D010938:Plant Oils; C049811:erucic acid; D000445:Aldehyde Oxidoreductases; C021351:hexadecanal dehydrogenase (acylating)