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Title

Characterization and partial purification of acyl-CoA:glycerol 3-phosphate acyltransferase from sunflower (Helianthus annuus L.) developing seeds

AuthorsRuiz-López, Noemí ; Garcés Mancheño, Rafael ; Harwood, John L.; Martínez-Force, Enrique
KeywordsAcyltransferase
Glycerol 3-phosphate
Hellianthus annuus
Microsomal membrane
Sunflower
Seeds
Issue DateFeb-2010
PublisherElsevier
CitationPlant Physiology and Biochemistry 48(2-3): 73- 80 (2010)
AbstractThe glycerol 3-phosphate acyltransferase (GPAT, EC 2.3.1.15) from sunflower (Helianthus annuus L.) microsomes has been characterised and partially purified. The in vitro determination of activity was optimized, and the maximum value for GPAT activity identified between 15 and 20 days after flowering. The apparent Michaelis-Menten Km for the glycerol 3-phosphate was 354 μM. The preferred substrates were palmitoyl-CoA = linoleoyl-CoA > oleoyl-CoA with the lowest activity using stearoyl-CoA. High solubilisation was achieved using 0.75% Tween80 and the solubilised GPAT was partially purified by ion-exchange chromatography using a Hi-Trap DEAE FF column, followed by gel filtration chromatography using a Superose 12 HR column. The fraction containing the GPAT activity was analysed by SDS-PAGE and contained a major band of 60.1 kDa. Finally, evidence is provided which shows the role of GPAT in the asymmetrical distribution, between positions sn-1 and sn-3, of saturated fatty acids in highly saturated sunflower triacylglycerols. This work provides background information on the sunflower endoplasmic reticulum GPAT which may prove valuable for future modification of oil deposition in this important crop. © 2009 Elsevier Masson SAS. All rights reserved.
URIhttp://hdl.handle.net/10261/54496
DOI10.1016/j.plaphy.2009.12.001
Identifiersdoi: 10.1016/j.plaphy.2009.12.001
issn: 0981-9428
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