Two novel thioesterases are key determinants of the bimodal distribution of acyl chain length of Cuphea palustris seed oil.
The seed oil of Cuphea palustris has an unusual fatty-acyl composition, whereby the principal fatty-acyl groups, myristate (64%) and caprylate (20%), differ by more than two methylenes. We have isolated two thioesterase (TE) cDNAs from C. palustris, encoding proteins designated Cp FatB1 and Cp FatB2, which, when expressed in Escherichia coli, have TE activities specific for 8:0/10:0- and 14:0/16:0-acyl carrier protein substrates, respectively. The specific activities of the recombinant affinity-purified enzymes indicate that Cp FatB2 is kinetically superior to Cp FatB1. This result is consistent with the predominance of 14:0 in the seed oil, despite apparently equal mRNA abundance of the two transcripts in the seed. In C. palustris the expression of both sequences is confined to the seed tissues. Based on these findings we propose that these two enzymes are major factors determining the bimodal chain-length composition of C. palustris oil. Analysis of the immature and mature seed oil by reverse-phase high-performance liquid chromatography confirmed that the principal triglycerides contain both 8:0 and 14:0. This result indicates that both fatty acids are synthesized at the same time and in the same cells at all developmental stages during oil deposition, suggesting that the two TEs act together in the same fatty acid synthesis system.
Dehesh
K
K
Oils Division, Calgene Inc., Davis, California 95616, USA.
Edwards
P
P
Hayes
T
T
Cranmer
A M
AM
Fillatti
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D002210:Caprylates / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005227:Fatty Acids / Q000096:biosynthesis*; D015723:Gene Library; D008969:Molecular Sequence Data; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D010938:Plant Oils / Q000737:chemistry*; D010940:Plant Proteins; D010944:Plants / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D011994:Recombinant Proteins / Q000378:metabolism; D012639:Seeds / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution; D014280:Triglycerides / Q000737:chemistry
D002210:Caprylates; D018076:DNA, Complementary; D005227:Fatty Acids; D009227:Myristic Acids; D010938:Plant Oils; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D011994:Recombinant Proteins; D014280:Triglycerides; D019814:Myristic Acid; C098243:FatB2 protein, Cuphea palustris; D013869:Thiolester Hydrolases