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五种树坚果油体的结构组成与稳定性OA北大核心CSTPCD

Structural Composition and Stability of Oil Bodies from 5 Tree Nuts

中文摘要英文摘要

[目的]在油料种子中,脂类物质主要以油体的形式存在.核桃、榛子等树坚果是重要的木本油料,通过比较 5 种树坚果油体在形态、组成和稳定性上的差异,同时探讨三者间的关系,为木本坚果油脂的利用提供新途径.[方法]以核桃、巴旦木、榛子、薄壳山核桃、松子5种树坚果为研究对象,利用透射电镜对种仁细胞内油体的形态和大小进行观察比较;采用SDS-PAGE技术对油体膜蛋白进行分离,切取蛋白条带进行LC-MS/MS质谱鉴定;利用脂质组学对油体膜磷脂和油体内部甘油三酯组成进行分析;基于室温储藏条件下油体的形态变化,结合测定过氧化值和硫代巴比妥酸值,对油体稳定性进行评价.[结果]5种树坚果种仁细胞内均充斥油体和蛋白体,油体具完整包膜,形态多呈非对称椭球形,最大平均粒径为薄壳山核桃油体(2.34±0.48)μm,最小为巴旦木油体(1.21±0.19)μm.油质蛋白(oleosin)是5种树坚果最主要的油体膜蛋白组分,占膜蛋白总量的 72.76%—84.15%;其分子量分布较窄,为 14.7-18.8 kDa,但不同坚果中oleosin亚型数量存在较大差异.钙调油体蛋白(caleosin)占膜蛋白总量的14.34%—24.96%,其在不同坚果中的分子量接近,为26.5-27.1 kDa,且5种坚果中均只鉴定出一种caleosin构型.固醇油体蛋白(steroleosin)仅在核桃、薄壳山核桃和松子中被鉴定出,占膜蛋白总量的 8.95%—12.89%.脂质组成方面,5 种树坚果油体内部的甘油三酯组成差异较大,但膜磷脂组成近似.磷脂酰丝氨酸(PS)和磷脂酰胆碱(PC)是最主要的膜磷脂组分,两者合占膜磷脂总量的 67.91%—75.06%.膜磷脂中不饱和脂肪酰基链比例高达69.64%—74.52%,且其中38.64%—45.15%的脂肪酰基均为油酸酰基.室温下榛子油体表现出最高的稳定性,储藏过程中,油体逐渐聚集并融合,同时伴随脂质氧化程度的升高,最终导致乳析现象的发生.[结论]5 种树坚果以薄壳山核桃油体粒径最大,榛子油体稳定性最高.不同树坚果油体具有近似的膜蛋白和膜磷脂组成,但在oleosin异构体数量、油体内甘油三酯组成以及组分间比例上差异明显.其中,油体内脂质含量与oleosin的比例、油体膜磷脂与油体膜蛋白的比例可能对油体大小和稳定性具有重要影响.

[Objective]In oilseeds,lipids are mainly stored in the organelles called oil bodies.Tree nuts,such as walnut and hazelnut,were important oilseed crops.Among the 5 selected tree nuts,the differences in morphology,structure and stability of their oil bodies were evaluated and discussed,which could provide the new ideas for the utilization of the nut oils.[Method]5 tree nuts were used as materials,including walnut,almond,hazelnut,pecan,and pine nut.The morphology and size of the oil bodies inside the cells of different nut seeds were observed and compared by employing transmission electron microscope.The oil body membrane proteins were separated by SDS-PAGE,followed by the LC-MS/MS identification of the protein bands.The oil body membrane phospholipids,as well as the intrinsic triacylglycerols,were also analyzed through lipidomics.The stability of oil bodies was evaluated by analyzing their changes in morphology peroxide values and thiobarbituric acid values during storage under room temperature.[Result]The cells of the nut seeds were occupied by the oil bodies and proteosomes.The asymmetric spheroid-shaped oil bodies were covered by intact membranes.The pecan oil body had the largest average diameter(2.34±0.48)μm,while the almond oil body had the minimum(1.21±0.19)μm.For all nut samples,oleosin was the most important oil body membrane protein,accounting for 72.76%-84.15%of the total amount of membrane proteins.Oleosin had a narrow molecular weight distribution of 14.7-18.8 kDa,yet the numbers of oleosin isomers were quite different.Caleosin accounted for 14.34%-4.96%of the total amount of membrane proteins,and its molecular weight in different nuts was approximately 26.5-27.1 kDa.No caleosin isomer was found in each nut sample.Steroleosin,accounted for 8.95%-12.89%of the total amount of membrane proteins,was identified only in walnut,pecan and pine nut oil bodies.The oil bodies of the 5 tree nuts had different triacylglycerol compositions but similar membrane phospholipid compositions.Phospholipidine(PS)and phosphatidylcholine(PC)were the most important oil body membrane phospholipids,accounting for 67.91%-75.06%of the total amount of membrane phospholipids.The proportion of unsaturated fatty acyl chains in membrane phospholipids was 69.64%-74.52%,while 38.64%-45.15%of the fatty acyl groups were oleoyls.The hazelnut oil body showed the highest stability at room temperature.During storage,the oil bodies aggregated and fused gradually,accompanied by an increasing degree of lipid oxidation.Eventually the phase separation phenomenon in oil body emulsions was observed.[Conclusion]Among the oil bodies from 5 tree nuts,the pecan oil body was the largest in size,while the hazelnut oil body was the most stable.The different tree nut oil bodies had similar constituents of membrane proteins and membrane phospholipids,but there were significant differences in the number of oleosin isomers,triacylglycerol compositions and constituent proportions.The ratio of lipid content to oleosin,as well as the ratio of oil body membrane phospholipids to oil body membrane proteins,might have important influences on the size and stability of the oil bodies.

孙晓岩;金凤;杨旭畅;王丰俊;周晔

北京林业大学生物科学与技术学院/林木资源高效生产全国重点实验室/林业食品加工与安全北京市重点实验室,北京 100083中国林业科学研究院林业研究所/林木遗传育种全国重点实验室/国家林业和草原局林木培育重点实验室,北京 100091

树坚果;油体;油质蛋白;膜磷脂;稳定性

tree nuts;oil body;oleosin;membrane phospholipid;stability

《中国农业科学》 2024 (010)

2023-2034,中插45-中插48 / 16

国家自然科学基金(31901350)

10.3864/j.issn.0578-1752.2024.10.013

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