Physicochemical Characterisation of Sunflower Seed Oil Bodies Ex- Vivo

Oilseeds store energy as triacylglycerides during periods of dormancy in preparation for germination and the early stages of development. The triacylglyceride is stored in discrete organelles termed oil bodies. Oil bodies are formed during the synthesis of neutral lipids within the bilayer of cellular endoplasmic reticulum (ER); as lipid is synthesised it forms droplets of oil that swell distending the ER membrane and at a critical diameter separate from the ER by vesiculation forming independent organelles. These organelles are structurally stabilised by a phospholipid monolayer originating from the ER and the addition of highly amphiphilic oleosin proteins. Oil bodies have been shown previously to be extremely stable organelles that can be easily extracted and purified from oilseeds; our aim was to develop an understanding of the physical and chemical properties of sunflower oil bodies ex-vivo prior to their subsequent use in commercial products. Several novel findings were elucidated through this work: oil body phytochemical composition, their physical and oxidative stability and their ability to store and deliver flavour compounds. It was hypothesised that tocopherol is tightly associated with sunflower oil bodies. This was tested by recovering oil bodies from sunflower seed and washing them to remove extraneous proteins and associated phenolic compounds. Tocopherol remained with washed oil bodies (392 mg tocopherol.kg-1 oil body oil) and this population of tocopherol represented 38% of the total seed tocopherol. It was hypothesised that this high tocopherol concentration and its intrinsic association to oil body structures would contribute to an increased level of oxidative stability. Sunflower seed lipids were significantly more resistant to thermally induced oxidation when stabilised in oil body suspensions compared to sunflower oil emulsions stabilised by a range of commercial emulsifiers (sodium dodecyl sulfate, polyoxyethylenesorbitan monolaurate (tween 20) and dodecyltrimethylammonium bromide). Oxidative stability was assessed through lipid hydroperoxide concentration and the concentration of headspace hexanal. Maximum lipid hydroperoxide concentration in surfactant stabilised emulsions after 8 days at 45oC ranged between 26 and 333 mmol lipid hydroperoxide.kg-1 oil whereas lipid hydroperoxide concentrations in oil body suspensions did not exceed 12 mmol lipid hydroperoxide.kg-1 oil. In addition there was no development in oxidative rancidity over the 8 day storage trial of oil bodies stored at 5oC. The composition of phospholipids in a range of oil body preparations was assessed; purified oil bodies contained principally phosphatidylcholine (91%) and a smaller fraction of phosphatidylethanolamine (9%). Less purified preparations contained other phospholipid species; the presence of which was explained by contamination with either non-intrinsic cellular phospholipids or phospholipase D that catalysed the breakdown of phospholipids to phosphatidic acid. Mechanisms and the extent of oil body physical stability were assessed using charge analysis and resistance of oil body preparations to changes in temperature and pH. Oil bodies are stabilised by a combination of steric hindrance and electrostatic repulsion provided by the surface proteins and phospholipids. Oil bodies had a zeta potential of -30mV at neutral pH and the surface charge was pH dependant with an apparent isoelectric point of between pH3 and pH6 was calculated from electrophoretic mobility, streaming potential and creaming stability measurements. Purified oil bodies were physically stable to thermal stresses up to 45oC for 2 days, although less purified preparations lost structural integrity at temperatures above 25oC. When assessed for their ability to delivery flavour molecules, oil bodies had comparable bulk phase properties to artificial emulsions stabilised by tween 20. Oil bodies did show a greater rate of flavour delivery during headspace dilution, when compared with the model artificial emulsions, suggesting commercial benefits may be gained through the incorporation of oil bodies into commercial emulsions. The key findings of this work are that oil bodies are extremely stable organelles that are resistant to thermal stress and physical processing. When lipid is stored within oil bodies it has greater resistance to the onset of lipid oxidation which may be explained by the intrinsic association of phospholipids, proteins and phytochemicals (vitamin E).