Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1990 Nov 15;272(1):31–38. doi: 10.1042/bj2720031

Regulation of triacylglycerol biosynthesis in embryos and microsomal preparations from the developing seeds of Cuphea lanceolata.

M Bafor 1, L Jonsson 1, A K Stobart 1, S Stymne 1
PMCID: PMC1149652  PMID: 2264835

Abstract

Embryos of Cuphea lanceolata have more than 80 mol% of decanoic acid ('capric acid') in their triacylglycerols, while this fatty acid is virtually absent in phosphatidylcholine (PtdCho). Seed development was complete 25-27 days after pollination, with rapid triacylglycerol deposition occurring between 9 and 24 days. PtdCho amounts increased until day 15 after pollination. Analysis of embryo lipids showed that the diacylglycerol (DAG) pool consisted of mainly long-chain molecular species, with a very small amount of mixed medium-chain/long-chain glycerols. Almost 100% of the fatty acid at position sn-2 in triacylglycerols (TAG) was decanoic acid. When equimolar mixtures of [14C]decanoic and [14C]oleic acid were fed to whole detached embryos, over half of the radioactivity in the DAG resided in [14C]oleate, whereas [14C]decanoic acid accounted for 93% of the label in the TAG. Microsomal preparations from developing embryos at the mid-stage of TAG accumulation catalysed the acylation of [14C]glycerol 3-phosphate with either decanoyl-CoA or oleoyl-CoA, resulting in the formation of phosphatidic acid (PtdOH), DAG and TAG. Very little [14C]glycerol entered PtdCho. In combined incubations, with an equimolar supply of [14C]oleoyl-CoA and [14C]decanoyl-CoA in the presence of glycerol 3-phosphate, the synthesized PtdCho species consisted to 95% of didecanoic and dioleic species. The didecanoyl-glycerols were very selectively utilized over the dioleoylglycerols in the production of TAG. Substantial amounts of [14C]oleate, but not [14C]decanoate, entered PtdCho. The microsomal preparations of developing embryos were used to assess the acyl specificities of the acyl-CoA:sn-glycerol-3-phosphate acyltransferase (GPAT, EC 2.3.1.15) and the acyl-CoA:sn-1-acyl-glycerol-3-phosphate acyltransferase (LPAAT, EC 2.3.1.51) in Cuphea lanceolata embryos. The efficiency of acyl-CoA utilization by the GPAT was in the order decanoyl = dodecanoyl greater than linoleoyl greater than myristoyl = oleoyl greater than palmitoyl. Decanoyl-CoA was the only acyl donor to be utilized to any extent by the LPAAT when sn-decanoylglycerol 3-phosphate was the acyl acceptor. sn-1-Acylglycerol 3-phosphates with acyl groups shorter than 16 carbon atoms did not serve as acyl acceptors for long-chain (greater than or equal to 16 carbon atoms) acyl-CoA species. On the basis of the results obtained, we propose a schematic model for triacylglycerol assembly and PtdCho synthesis in a tissue specialized in the synthesis of high amounts of medium-chain fatty acids.

Full text

PDF
33

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Cao Y. Z., Huang A. H. Acyl coenzyme a preference of diacylglycerol acyltransferase from the maturing seeds of cuphea, maize, rapeseed, and canola. Plant Physiol. 1987 Jul;84(3):762–765. doi: 10.1104/pp.84.3.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Huang K. P., Stumpf P. K. Fat metabolism in higher plants. XLIV. Fatty acid synthesis by a soluble fatty acid synthetase from Sclanum tuberosum. Arch Biochem Biophys. 1971 Apr;143(2):412–427. doi: 10.1016/0003-9861(71)90228-1. [DOI] [PubMed] [Google Scholar]
  4. Ichihara K., Asahi T., Fujii S. 1-Acyl-sn-glycerol-3-phosphate acyltransferase in maturing safflower seeds and its contribution to the non-random fatty acid distribution of triacylglycerol. Eur J Biochem. 1987 Sep 1;167(2):339–347. doi: 10.1111/j.1432-1033.1987.tb13342.x. [DOI] [PubMed] [Google Scholar]
  5. Ichihara K. sn-Glycerol-3-phosphate acyltransferase in a particulate fraction from maturing safflower seeds. Arch Biochem Biophys. 1984 Aug 1;232(2):685–698. doi: 10.1016/0003-9861(84)90589-7. [DOI] [PubMed] [Google Scholar]
  6. KATES M. SIMPLIFIED PROCEDURES FOR HYDROLYSIS OR METHANOLYSIS OF LIPIDS. J Lipid Res. 1964 Jan;5:132–135. [PubMed] [Google Scholar]
  7. KENNEDY E. P. Biosynthesis of complex lipids. Fed Proc. 1961 Dec;20:934–940. [PubMed] [Google Scholar]
  8. Kanda P., Wells M. A. Facile acylation of glycerophosphocholine catalyzed by trifluoroacetic anhydride. J Lipid Res. 1981 Jul;22(5):877–879. [PubMed] [Google Scholar]
  9. LANDS W. E., HART P. METABOLISM OF GLYCEROLIPIDS. VI. SPECIFICITIES OF ACYL COENZYME A: PHOSPHOLIPID ACYLTRANSFERASES. J Biol Chem. 1965 May;240:1905–1911. [PubMed] [Google Scholar]
  10. Ohlrogge J. B., Kuhn D. N., Stumpf P. K. Subcellular localization of acyl carrier protein in leaf protoplasts of Spinacia oleracea. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1194–1198. doi: 10.1073/pnas.76.3.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Oo K. C., Huang A. H. Lysophosphatidate acyltransferase activities in the microsomes from palm endosperm, maize scutellum, and rapeseed cotyledon of maturing seeds. Plant Physiol. 1989 Dec;91(4):1288–1295. doi: 10.1104/pp.91.4.1288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Shimakata T., Stumpf P. K. The purification and function of acetyl coenzyme A:acyl carrier protein transacylase. J Biol Chem. 1983 Mar 25;258(6):3592–3598. [PubMed] [Google Scholar]
  13. Slack C. R., Roughan P. G., Browse J. Evidence for an oleoyl phosphatidylcholine desaturase in microsomal preparations from cotyledons of safflower (Carthamus tinctorius) seed. Biochem J. 1979 Jun 1;179(3):649–656. doi: 10.1042/bj1790649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stobart A. K., Stymne S. The interconversion of diacylglycerol and phosphatidylcholine during triacylglycerol production in microsomal preparations of developing cotyledons of safflower (Carthamus tinctorius L.). Biochem J. 1985 Nov 15;232(1):217–221. doi: 10.1042/bj2320217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stymne S., Appelqvist L. A. The biosynthesis of linoleate from oleoyl-CoA via oleoyl-phosphatidylcholine in microsomes of developing safflower seeds. Eur J Biochem. 1978 Oct;90(2):223–229. doi: 10.1111/j.1432-1033.1978.tb12594.x. [DOI] [PubMed] [Google Scholar]
  16. Sánchez M., Nicholls D. G., Brindley D. N. [The relationship between palmitoyl-coenzyme A synthetase activity and esterification of sn-glycerol 3-phosphate in rat liver mitochondria]. Biochem J. 1973 Apr;132(4):697–706. doi: 10.1042/bj1320697. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES