Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (10): 2023-2034.doi: 10.3864/j.issn.0578-1752.2024.10.013

• FOOD SCIENCE AND ENGINEERING • Previous Articles     Next Articles

Structural Composition and Stability of Oil Bodies from 5 Tree Nuts

SUN XiaoYan1(), JIN Feng1, YANG XuChang1, WANG FengJun1(), ZHOU Ye2()   

  1. 1 Beijing Key Laboratory of Forestry Food Processing and Safety/State Key Laboratory of Efficient Production of Forest Resources/College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083
    2 Research Institute of Forestry, Chinese Academy of Forestry/State Key Laboratory of Tree Genetics and Breeding/Key Laboratory of Tree Breeding and Cultivation of the State Forestry and Grassland Administration, Beijing 100091
  • Received:2023-12-28 Accepted:2024-03-01 Online:2024-05-16 Published:2024-05-23
  • Contact: WANG FengJun, ZHOU Ye

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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

Fig. 1

Ultrastructure of oil bodies of the 5 tree nuts CW: Cell wall; OB: Oil body; PB: Protein body. A: Walnut; B: Almond; C: Hazelnut; D: Pecan; E: Pine (A-E×2900)"

Table 1

The average number, average particle size and particle size distribution of oil bodies of the 5 tree nuts"

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 (%)
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

Fig. 2

SDS-PAGE electrophoresis analysis of the oil body proteins of the 5 tree nuts M: Protein molecular marker; A: Pine; B: Pecan; C: Hazelnut; D: Almond; E: Walnut"

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 3

Compositions and contents of the phospholipids in oil bodies of the 5 tree nuts"

序号
No.		 磷脂种类
Phospholipid species		 含量Content (nmol∙g-1)
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

Fig. 3

Distribution of the main fatty acids in oil body membrane phospholipids of the 5 tree nuts A: Walnut; B: Almond; C: Hazelnut; D: Pecan; E: Pine"

Fig. 4

Stability of the oil bodies of the 5 tree nuts A: Walnut; B: Almond; C: Hazelnut; D: Pecan; E: Pine"

Fig. 5

Changes of the peroxide values and thiobarbituric acid values of the oil bodies during storage Different lowercase letters at the same time indicate significant difference (P<0.05)"

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