Microsomal membrane preparations from the developing endosperm of castor bean (Ricinus communis) catalysed the transfer of oleate from [14C]oleoyl-CoA to phosphatidylcholine (PtdCho). In the presence of NADH, radioactive ricinoleate (12-hydroxyoctadec-9-enoate) was synthesized from [14C]oleate, and this was largely recovered in PtdCho and as free fatty acid. The addition of unlabelled ricinoleoyl-CoA to these incubation mixtures did not increase the low [14C]ricinoleate concentration found in the acyl-CoA fraction nor decrease the [14C]ricinoleate concentration in PtdCho and free fatty acid, and thus no evidence was obtained for a hydroxylation with oleoyl-CoA as a substrate. The addition of NADH, necessary for the formation of ricinoleate, caused a decrease of the total radioactivity in PtdCho with a corresponding increase in the amount of label in free ricinoleic acid. This increase was due to the action of a phospholipase A, which released ricinoleic acid but not oleic acid from PtdCho. Such a phospholipase activity, attacking ricinoleoyl-PtdCho but not oleoyl-PtdCho, was also demonstrated in microsomal preparations from developing cotyledons of safflower and oil-seed rape. An analysis of the acyl groups at different positions in microsomal PtdCho of castor bean showed that ricinoleate was almost entirely associated with position sn-2. Likewise the [14C]ricinoleate in [14C]PtdCho formed after incubations with microsomal preparations with NADH and [14C]oleoyl-CoA resided in position sn-2 with none in position sn-1. In contrast, the [14C]linoleate formed by desaturation of [14C]oleoyl-PtdCho was present at both positions. In the presence of ATP, CoA and Mg2+, the ricinoleate acid released from PtdCho was activated to ricinoleoyl-CoA. The ricinoleoyl-CoA was an efficient acyl donor in the acylation of glycerol 3-phosphate (Gro3P) to yield phosphatidic acid and triacylglycerols. In microsomal preparations incubated with an equimolar mixture of [14C]oleoyl-CoA and [14C]ricinoleoyl-CoA in the presence of Gro3P, only a minor amount of [14C]ricinoleate entered PtdCho, and this was believed to be via the exchange of phosphocholine groups between a diacylglycerol pool and the PtdCho. On the basis of our results, a scheme of ricinoleate formation and its incorporation into triacylglycerols in castor-bean endosperm is proposed.
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December 1991
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Research Article|
December 01 1991
Ricinoleic acid biosynthesis and triacylglycerol assembly in microsomal preparations from developing castor-bean (Ricinus communis) endosperm
M Bafor;
M Bafor
1Department of Plant Physiology, Swedish University of Agricultural Sciences, P.O. Box 7047, S-750 07 Uppsala, Sweden
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M A Smith;
M A Smith
1Department of Plant Physiology, Swedish University of Agricultural Sciences, P.O. Box 7047, S-750 07 Uppsala, Sweden
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L Jonsson;
L Jonsson
1Department of Plant Physiology, Swedish University of Agricultural Sciences, P.O. Box 7047, S-750 07 Uppsala, Sweden
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K Stobart;
K Stobart
1Department of Plant Physiology, Swedish University of Agricultural Sciences, P.O. Box 7047, S-750 07 Uppsala, Sweden
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S Stymne
S Stymne
1Department of Plant Physiology, Swedish University of Agricultural Sciences, P.O. Box 7047, S-750 07 Uppsala, Sweden
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Biochem J (1991) 280 (2): 507–514.
Citation
M Bafor, M A Smith, L Jonsson, K Stobart, S Stymne; Ricinoleic acid biosynthesis and triacylglycerol assembly in microsomal preparations from developing castor-bean (Ricinus communis) endosperm. Biochem J 1 December 1991; 280 (2): 507–514. doi: https://doi.org/10.1042/bj2800507
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