Characterization of acyl-ACP thioesterases of mangosteen (Garcinia mangostana) seed and high levels of stearate production in transgenic canola.
Acyl-acyl-carrier protein (ACP) thioesterases are, at least in part, responsible for the fatty acyl chain length composition of seed storage oils. Acyl-ACP thioesterases with specificity for each of the saturated acyl-ACP substrates from 8:0 through 16:0 have been cloned, with the exception of 18:0, and are members of the FatB class of thioesterases. The authors have determined that the tropical tree species mangosteen (Garcinia mangostana) stores 18:0 (stearate) in its seed oil in amounts of up to 56% by weight. Acyl-ACP thioesterase activity as measured in crude mangosteen seed extracts showed a preference for 18:1-ACP substrates, but had significant activity with 18:0 relative to that with 16:0-ACP, suggesting a thioesterase might be involved in the production of stearate. Three distinct acyl-ACP thioesterases were cloned from mangosteen seed cDNA; two representative of the FatA class and one representative of the FatB class. When expressed in vitro, the enzyme encoded by one of the FatAs (Garm FatA1) while preferring 18:1-ACP showed relatively low activity with 16:0-ACP as compared to 18:0-ACP, similar to the substrate preferences shown by the crude seed extract. Expression of Garm FatA1 in Brassica seeds led to the accumulation of stearate up to 22% in seed oil. These results suggest that Garm FatA1 is at least partially responsible for determining the high stearate composition of mangosteen seed oil and that FatA as well FatB thioesterases have evolved for specialized roles.
Calgene, Inc., Davis, CA 95616, USA.
D J
DJ
Hawkins
J C
JC
Kridl
D000595:Amino Acid Sequence; D001937:Brassica / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005638:Fruit / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010938:Plant Oils / Q000737:chemistry; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012639:Seeds / Q000737:chemistry / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D013229:Stearic Acids / Q000378:metabolism*; D013379:Substrate Specificity; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000378:metabolism*
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D010938:Plant Oils; D013229:Stearic Acids; C031183:stearic acid; D013869:Thiolester Hydrolases; C018748:oleoyl-(acyl-carrier-protein) hydrolase