Investigation of diacylglycerol acyltransferase (DGAT) activity of microsomes from the seeds of three euphorbs |
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| Authors: | MJ Pascual-Villalobos MD López MT Castells MD Alcázar |
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| Institution: | 1. The Key Laboratory of Plant Resources and Chemistry of Arid Zone, Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Urumqi 830011, China;2. State Key Laboratory Basis of Xinjiang Indigenous Medicinal Plants Resource Utilization, Urumqi 830011, China;3. University of the Chinese Academy of Sciences, Beijing 100039, China;1. State Key Laboratory of Drug Research, Shanghai Institute of Meteria Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201201, People''s Republic of China;2. University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, People''s Republic of China;1. Department of Food Science and Technology, Keimyung University, Daegu 42601, Republic of Korea;2. The center for Traditional Microorganism Resource (TMR), Keimyung University, Daegu 42601, Republic of Korea;3. Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX 77843-4466, USA |
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| Abstract: | Unusual fatty acids such as ricinoleate (12-hydroxyoleic acid) occurring in Ricinus communis L. or vernoleate (12,13-epoxyoleic acid) occurring in Euphorbia lagascae L. are suitable for industrial uses. Euphorbia lathyris L. is also a potential new oilseed crop on account of its high content of oleic acid in the seeds. The objective of this work was to test in vitro the preferences of E. lathyris microsomes for its native substrates (oleoyl-CoA and diolein) and to compare with R. communis and E. lagascae systems.The diacylglycerol acyltransferase (DGAT) catalyses the final step in transferring a fatty acid moiety to a diacylglycerol (DAG) into a triacylglycerol (TAG). To study the DGAT activity in microsomes of the three euphorbs: (1) plants of the three species were grown in a glasshouse at Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (Murcia, Spain), (2) endosperms were removed from developing seeds and the tissue was extracted, (3) in vitro DGAT assays using 14C]-oleoyl-CoA with or without 1,2-diolein were carried out and 4) labelled TAG were recorded using a molecular imager and a scintillation counter.Incorporation of 14C]-oleoyl into TAG was greater in R. communis and E. lathyris (72–89% of total TAG) than in E. lagascae. Adding exogenous 1,2-diolein (1 mM) to E. lagascae microsomes increased the amount of labelled TAG to 39%, suggesting that other acyl groups were being incorporated as well. R. communis and E. lagascae microsomes gave more-polar radiolabelled TAGs than E. lathyris possibly because endogenous DAGs (not 1,2-diolein) were being used in the reaction. Although E. lathyris microsomes showed specificity towards 1,2-diolein as a substrate, the preparations from R. communis, E. lagascae and E. lathyris were able to use the acyl donor and acyl receptor, possibly suggesting that DGAT enzymes would not be a limiting factor to engineer Euphorbiaceae crops with functionalized fatty acids. |
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