五种树坚果油体的结构组成与稳定性

doi:10.3864/j.issn.0578-1752.2024.10.013

Abstract

Abstract:

【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.

Key words: tree nuts, oil body, oleosin, membrane phospholipid, stability

SUN XiaoYan, JIN Feng, YANG XuChang, WANG FengJun, ZHOU Ye. Structural Composition and Stability of Oil Bodies from 5 Tree Nuts[J].Scientia Agricultura Sinica, 2024, 57(10): 2023-2034.

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Figures/Tables 8

Fig. 1

Table 1

Fig. 2

Table 2

Identification of the oil body proteins of the 5 tree nuts by LC-MS/MS"

ID号 Accession ID	鉴定蛋白名称[物种] Description [Species]	分子量 MW (kDa)	等电点 Calc. pI	蛋白匹配度打分 Score sequest	特征肽段数Unique peptides
油质蛋白 Oleosin
A0A2I4GES3	油质蛋白5-like[核桃] Oleosin 5-like [Walnut]	16.3	10.08	494.3	10
A0A2I4G1C3	油质蛋白18.2 kDa -like[核桃] Oleosin 18.2 kDa-like [Walnut]	16.6	9.91	461.39	13
G8H6H9	油质蛋白[核桃] Oleosin [Walnut]	14.7	10.14	207.35	4
A0A2I4GSQ3	油质蛋白-like[核桃] Oleosin-like [Walnut]	16.8	8.88	180.41	9
G8H6H8	油质蛋白[核桃] Oleosin [Walnut]	14.8	9.63	143.06	4
A0A2I4DRG3	油质蛋白18.2 kDa -like[核桃] Oleosin 18.2 kDa-like [Walnut]	15.9	9.72	141.34	6
A0A2I4GNJ1	油质蛋白[核桃] Oleosin [Walnut]	15.6	9.54	79.8	4
A0A5E4ET55	油质蛋白[巴旦木] Oleosin [Almond]	16.6	9.98	598.45	12
Q43804	油质蛋白1[巴旦木] Oleosin 1 [Almond]	15.6	9.63	497.89	7
A0A5E4FAC4	油质蛋白[巴旦木] Oleosin [Almond]	13.1	9.41	36.41	1
A0A5E4ET40	预测：油质蛋白[巴旦木] Predicted: oleosin [Almond]	14.3	6.52	26.59	1
A0A5E4F6M0	预测：油质蛋白[巴旦木] Predicted: oleosin [Almond]	18.8	9.92	106.02	5
A0A5E4GEY2	油质蛋白[巴旦木] Oleosin [Almond]	24	7.68	254.67	10
Q84T91	油质蛋白 Cor a 13[榛子] Oleosin Cor a 13 [Hazelnut]	14.7	9.98	318.25	6
Q84T21	油质蛋白 Cor a 12[榛子] Oleosin Cor a 12 [Hazelnut]	16.7	10.54	278.34	8
C0HM28	油质蛋白 Cor a 15[榛子] Oleosin Cor a 15 [Hazelnut]	17.7	9.54	243.87	10
A0A8T1PXN9	油质蛋白[薄壳山核桃] Oleosin [Pecan]	14.5	9.79	129.39	4
A0A8T1QQ11	油质蛋白[薄壳山核桃] Oleosin [Pecan]	14.6	10.35	160.65	5
A0A8T1PRE2	油质蛋白[薄壳山核桃] Oleosin [Pecan]	14.6	9.98	208.69	4
A0A8T1Q1F4	油质蛋白[薄壳山核桃] Oleosin [Pecan]	15.6	9.58	88.52	4
A0A8T1QWQ9	油质蛋白[薄壳山核桃] Oleosin [Pecan]	16.2	10.27	219.88	8
A0A8T1NSX3	油质蛋白[薄壳山核桃] Oleosin [Pecan]	16.2	10.35	46.45	5
A0A8T1NEK6	油质蛋白[薄壳山核桃] Oleosin [Pecan]	16.9	9.22	88.16	8
A0A8T1NXG5	油质蛋白[薄壳山核桃] Oleosin [Pecan]	17.5	9.92	265.47	13
A0A1I9R3Y6	油质蛋白[松子] Oleosin [Pine]	15.6	7.64	131.63	5
A0A060L4I9	油质蛋白L[松子] Oleosin L [Pine]	15.1	8.44	5.61	1
钙调油体蛋白 Caleosin
A0A2I4DSD4	过氧合酶-like[核桃] Peroxygenase-like [Walnut]	26.9	5.87	682.98	21
A0A5E4EKE0	预测：过氧合酶[巴旦木] Predicted: peroxygenase [Almond]	26.5	6.05	374.26	12
A0A6C0PBB1	油体钙调蛋白1[榛子] Caleosin 1 [Hazelnut]	26.8	6.05	395.81	18
A0A8T1RPB9	过氧合酶[薄壳山核桃] Peroxygenase [Pecan]	27.1	6.3	548.93	20
A0A1I9R3Y5	过氧合酶[松子] Peroxygenase [Pine]	26.5	6.44	155.52	5
固醇油体蛋白 Steroleosin
A0A833X2K3	11-β-羟基类固醇脱氢酶-like 5[核桃] 11-beta-hydroxysteroid dehydrogenase -like 5 [Walnut]	46.1	6.46	487.34	19
A0A8T1QPV6	11-β-羟基类固醇脱氢酶-like 5[薄壳山核桃] 11-beta-hydroxysteroid dehydrogenase-like 5 [Pecan]	40.9	6.33	500.24	20
Q0GJJ3	11-β-羟基类固醇脱氢酶[松子] 11-beta-hydroxysteroid dehydrogenase-like [Pine]	37.7	9.44	4.96	1
A0A140FAN3	11-β-羟基类固醇脱氢酶[松子] 11-beta-hydroxysteroid dehydrogenase [Pine]	40.7	8.16	244.03	11

Table 2

Table 3

Fig. 3

Fig. 4

Fig. 5

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序号 No.	物种 Species	平均个数 Average quantities	平均粒径 Average diameter (μm)	粒径分布Size distribution (%)
序号 No.	物种 Species	平均个数 Average quantities	平均粒径 Average diameter (μm)	0—1 μm	1—2 μm	2—2.5 μm	>2.5 μm
1	核桃 Walnut	250.48±20.57d	1.36±0.22d	40.28±3.32b	31.03±4.28d	9.58±3.57d	19.11±4,24d
2	巴旦木 Almond	367.93±17.95b	1.21±0.19e	28.34±4.59d	59.61±5.32a	12.05±3.12c	0.00±0.00e
3	榛子 Hazelnut	314.36±24.08c	1.42±0.25c	25.41±6.34e	32.24±8.34c	20.05±4.56a	22.30±6.02b
4	薄壳山核桃 Pecan	111.14±18.32e	2.34±0.48a	41.27±4.14a	25.13±4.67e	7.28±4.35e	26.32±3.09a
5	松子 Pine	430.64±45.01a	1.61±0.34b	30.23±4.67c	35.49±6.04b	12.98±5.67b	21.30±5.82c

序号 No.	磷脂种类 Phospholipid species	含量Content (nmol∙g^-1)
序号 No.	磷脂种类 Phospholipid species	核桃 Walnut	巴旦木 Almond	榛子 Hazelnut	薄壳山核桃 Pecan	松子 Pine
1	磷脂酰丝氨酸Phosphatidylserine (PS)	19.08±4.38c	17.51±2.99d	34.24±5.85a	25.38±2.01b	24.45±9.50b
2	磷脂酰胆碱Phosphatidylcholine (PC)	17.10±0.94d	27.37±2.34b	30.59±1.27a	21.31±1.16c	22.19±1.92c
3	磷脂酰甘油Phosphatidylglycerol (PG)	8.64±0.17d	9.41±0.21b	10.22±0.57a	9.48±0.60b	9.14±0.19c
4	磷脂酰肌醇Phosphatidyl inositol (PI)	2.96±0.39d	7.46±1.21a	5.35±0.77b	2.04±0.32e	3.99±0.40c
5	磷脂酰乙醇胺Phosphatidylethanolamine (PE)	1.52±0.30d	2.66±0.40b	4.00±0.69a	1.61±0.41d	2.03±0.23c
6	溶血磷脂酸Lysophosphatidic acid (LPA)	0.77±0.30c	0.61±0.14d	0.91±0.35b	1.43±0.09a	0.71±0.27cd
7	溶血磷脂酰胆碱Lysophosphatidylcholine (LPC)	0.45±0.04c	0.47±0.04fc	0.46±0.05c	0.97±0.07a	0.54±0.03b
8	溶血磷脂酰肌醇Lysophosphatidylinositol (LPI)	0.31±0.01cd	0.41±0.01a	0.37±0.02b	0.33±0.003c	0.27±0.01d
9	磷脂酸Phosphatidic acid (PA)	0.15±0.02b	0.15±0.03b	0.19±0.02a	0.15±0.02b	0.16±0.01b
10	磷脂酰甲醇Phosphatidylcarbinol (PmeOH)	0.032±0.001a	0.036±0.002a	0.036±0.002a	0.04±0.003a	0.03±0.001a

Structural Composition and Stability of Oil Bodies from 5 Tree Nuts

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