不同品种汉麻籽蛋白质结构与功能特性分析

宋炜昱; 尹浩; 钟宇; 王丹凤; 邓云

doi:10.13386/j.issn1002-0306.2022070122

不同品种汉麻籽蛋白质结构与功能特性分析

doi: 10.13386/j.issn1002-0306.2022070122

上海交通大学农业与生物学院食品科学与工程系，上海 200240

基金项目: 云南省科技厅科技创新引导与科技型企业培育计划（202204BP090031）

详细信息

作者简介:
宋炜昱（1998−），男，硕士研究生，研究方向：食品加工与营养，E-mail：thechosenonesjtu@sjtu.edu.cn

通讯作者:
邓云（1973−），男，博士，教授，研究方向：食品加工与营养，E-mail：y_deng@ sjtu.edu.cn

中图分类号: TS201.2
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出版历程
- 收稿日期: 2022-07-13
- 刊出日期: 2023-05-15

Analysis of Protein Structure and Functional Properties of Hemp Seeds of Different Varieties

Department of Food Science and Engineering, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China

摘要

摘要: 本文通过碱溶酸沉法提取了四个品种汉麻籽分离蛋白（Hemp protein isolate，HPI），测定了其分子量，并通过圆二色谱，游离巯基和二硫键含量以及表面疏水性表征其结构，最后测定了其功能特性并进行了功能特性和空间结构间的相关性分析。结果表明：HPI主要由含有两个亚基的麻仁球蛋白（约为50 kDa）和白蛋白（10~15 kDa）组成。四种HPI的二级结构中自由卷曲约占50%。东北的HPI二硫键含量为（5.20±0.32）μmol/g，拥有更稳定的结构；山西的HPI游离巯基为（15.18±0.32）μmol/g，更容易形成聚集体。在pH为4~6时，HPI的溶解度仅为4.85%~31.48%；其持水性和持油性分别为3.26~4.24和4.36~6.03 g/g，优于大部分植物蛋白；乳化能力和起泡能力较差。α-螺旋和表面疏水性与乳化性呈显著正相关性（r=0.70，0.65，P<0.05）；乳化稳定性与自由卷曲呈极显著正相关性（r=0.74，P<0.01），与二硫键呈极显著负相关性（r=−0.72，P<0.01）。综上，不同品种HPI的功能特性和空间结构有较大差异，研究结果可为汉麻籽分离蛋白的精深加工及其在不同食品及配方中的应用提供一定的理论依据。
- 汉麻籽蛋白 /
- 功能特性 /
- 圆二色谱 /
- 蛋白质结构 /
- 相关性分析
Abstract: In this paper, hemp protein isolate (HPI) of four different varieties was extracted by alkali dissolution and acid precipitation. The molecular weight of HPI was determined and its structures were characterized by circular dichroism, free sulfhydryl and disulfide bond content and surface hydrophobicity. Finally, its functional properties were determined and the correlation analysis between functional properties and spatial structure was conducted. The results showed that HPI was mainly composed of edestin (~50 kDa) and albumin (10~15 kDa), and the former had two subunits. Random coil accounted for about 50% of the secondary structure. The content of disulfide bond in HPI from Dongbei was (5.20±0.32) μmol/g which means a more stable structure, and the content of free sulfhydryl groups HPI from Shanxi was (15.18±0.32) μmol/g that means it was easier to form aggregates. At pH4~6, the solubility of HPI was only 4.85%~31.48%. The water holding capacity and oil holding capacity of HPI were 3.26~4.24 and 4.36~6.03 g/g, respectively, superior to most plant proteins. Compared with other plant proteins, HPI had poor emulsifying and foaming capacity. Alpha-helix and surface hydrophobicity were significantly positive in correlation with emulsifying ability (r=0.70, 0.65, P<0.05). There was highly significant positive correlation between emulsifying stability and random coil (r=0.74, P<0.01), and distinct negative correlation between emulsifying stability and disulfide bond (r=−0.72, P<0.01). Overall, the functional properties and spatial structure of different varieties of HPI were quite different. The research results could provide a theoretical basis for the intensive processing of HPI and its application in different foods and formulas.
- hemp protein isolate /
- functional properties /
- circular dichroism /
- protein structure /
- correlation analysis

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图 1 汉麻籽分离蛋白的凝胶电泳图

注：图中M表示标志物，D、G、N、S表示HPI来自东北、广西、内蒙古、山西。

Figure 1. SDS-PAGE of hemp protein isolate

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图 2 汉麻籽分离蛋白的二级结构

Figure 2. Secondary structure of hemp protein isolate

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图 3 汉麻籽分离蛋白的游离巯基和二硫键含量及表面疏水性

Figure 3. Free sulfhydryl, disulfide bond content and surface hydrophobicity of hemp protein isolate

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图 4 不同pH条件下汉麻籽分离蛋白的溶解度

Figure 4. Solubility of hemp protein isolate under different pH conditions

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图 5 汉麻籽分离蛋白的持水性和持油性

Figure 5. Water holding capacity and oil holding capacity of hemp protein isolate

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图 6 汉麻籽分离蛋白的乳化性能

Figure 6. Emulsifying properties of hemp protein isolate

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图 7 汉麻籽分离蛋白的起泡性能

注：ns表示无显著性差异，P>0.05。

Figure 7. Foaming properties of hemp protein isolate

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表 1 汉麻籽分离蛋白功能特性和结构之间的相关性分析

Table 1. Correlation analysis between functional properties and structure of hemp protein isolate

	功能特性						结构特性
	持水性	持油性	乳化性	乳化稳定性	起泡性	起泡稳定性	α-螺旋	β-折叠	自由卷曲	游离巯基	二硫键	表面疏水性
持水性	1.00	0.32	0.01	0.78^**	0.26	0.10	−0.05	−0.30	0.55	0.18	−0.38	−0.21
持油性		1.00	0.32	0.26	0.46	−0.31	−0.26	0.30	0.08	0.15	−0.17	0.76^**
乳化性			1.00	−0.42	0.31	0.30	0.701^*	−0.58^*	−0.54	−.086^**	0.76^**	0.65^*
乳化稳定性				1.00	0.23	−0.06	−0.44	0.10	0.74^**	0.60^*	−0.72^**	−0.29
起泡性					1.00	−0.28	0.27	−0.25	−0.16	−0.14	0.09	0.39
起泡稳定性						1.00	0.38	−0.33	−0.26	−0.39	0.38	−0.15
α-螺旋							1.00	−0.88^**	−0.70^*	−0.93^**	0.89^**	0.07
β-折叠								1.00	0.25	0.75^**	−0.61^*	0.19
自由卷曲									1.00	0.74^**	−0.89^**	−0.42
游离巯基										1.00	−0.96^**	−0.31
二硫键											1.00	0.32
表面疏水性												1.00
注：显著性水平， P<0.05；*，P<0.01。

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参考文献(32)

[1]	ANDRE C M, HAUSMAN J F, GUERRIERO G. Cannabis sativa: The plant of the thousand and one molecules[J]. Frontiers in Plant Science,2016,7:19.
[2]	SHEN P, GAO Z, FANG B, et al. Ferreting out the secrets of industrial hemp protein as emerging functional food ingredients[J]. Trends in Food Science & Technology,2021,112:1−15.
[3]	TANG K L, WANG J Y, YANG Y, et al. Fiber hemp (Cannabis sativa L.) yield and its response to fertilization and planting density in China[J]. Industrial Crops & Products,2022,177:114542.
[4]	FRASSINETTI S, MOCCIA E, CALTAVUTURO L, et al. Nutraceutical potential of hemp (Cannabis sativa L.) seeds and sprouts[J]. Food Chemistry,2018,262:56−66. doi: 10.1016/j.foodchem.2018.04.078
[5]	TEH S S, BEKHIT A E A, CARNE A, et al. Antioxidant and ACE-inhibitory activities of hemp (Cannabis sativa L.) protein hydrolysates produced by the proteases AFP, HT, Pro-G, actinidin and zingibain[J]. Food Chemistry,2016,203:199−206. doi: 10.1016/j.foodchem.2016.02.057
[6]	NONGONIERMA A B, FITZGERALD R J. Investigation of the potential of hemp, pea, rice and soy protein hydrolysates as a source of dipeptidyl peptidase IV (DPP-IV) inhibitory peptides[J]. Food Digestion: Research and Current Opinion,2015,6:1−3. doi: 10.1007/s13228-015-0042-7
[7]	YU Z, WU S, ZHAO W, et al. Anti-alzheimers activity and molecular mechanism of albumin-derived peptides against AChE and BChE[J]. Food & Function,2018,9(2):1173−1178.
[8]	KING J W. The relationship between cannabis/hemp use in foods and processing methodology[J]. Current Opinion in Food Science,2019,28:32−40. doi: 10.1016/j.cofs.2019.04.007
[9]	予辑. 药食同源原料目录(2017版)[J]. 口腔护理用品工业,2017,27(6):24−28. [YU J. Medicinal and food ingredients directory (2017 version)[J]. Oral Care Industry,2017,27(6):24−28.
[10]	CHUAN-HE T, ZI T, XIAN-SHENG W, et al. Physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate[J]. Journal of Agricultural and Food Chemistry,2006,54(23):8945−8950. doi: 10.1021/jf0619176
[11]	WANG X S, TANG C H, YANG X Q, et al. Characterization, amino acid composition and in vitro digestibility of hemp (Cannabis sativa L.) proteins[J]. Food Chemistry,2008,107(1):11−18. doi: 10.1016/j.foodchem.2007.06.064
[12]	WANG Q, JIANG J, XIONG Y L. High pressure homogenization combined with pH shift treatment: A process to produce physically and oxidatively stable hemp milk[J]. Food Research International,2018,106:487−494. doi: 10.1016/j.foodres.2018.01.021
[13]	MALOMO S A, ALUKO R E. A comparative study of the structural and functional properties of isolated hemp seed (Cannabis sativa L.) albumin and globulin fractions[J]. Food Hydrocolloids,2015,43:743−752. doi: 10.1016/j.foodhyd.2014.08.001
[14]	何锦风, 陈天鹏, 卢蓉蓉, 等. 汉麻籽的综合利用及产业化研究[J]. 中国食品学报,2010,10(3):98−112. [HE J F, CHEN T P, LU R R, et al. Study on the comprehensive utilization and industrialization of hempseed (Fructus cannabis)[J]. Journal of Chinese Institute of Food Science and Technology,2010,10(3):98−112. doi: 10.3969/j.issn.1009-7848.2010.03.016
[15]	VONAPARTIS E, AUBIN M P, SEGUIN P, et al. Seed composition of ten industrial hemp cultivars approved for production in Canada[J]. Journal of Food Composition and Analysis,2015,39:8−12. doi: 10.1016/j.jfca.2014.11.004
[16]	CORRADO G, PANNICO A, ZARRELLI A, et al. Macro and trace element mineral composition of six hemp varieties grown as microgreens[J]. Journal of Food Composition and Analysis,2022,114:104750. doi: 10.1016/j.jfca.2022.104750
[17]	JIANG Y, ZHOU X, ZHENG Y, et al. Impact of ultrasonication/shear emulsifying/microwave-assisted enzymatic extraction on rheological, structural, and functional properties of Akebia trifoliata (Thunb.) Koidz. seed protein isolates[J]. Food Hydrocolloids,2021,112:106355. doi: 10.1016/j.foodhyd.2020.106355
[18]	NASROLLAHZADEH F, ROMAN L, SWARAJ V J S, et al. Hemp (Cannabis sativa L.) protein concentrates from wet and dry industrial fractionation: Molecular properties, nutritional composition, and anisotropic structuring[J]. Food Hydrocolloids,2022,131:107755. doi: 10.1016/j.foodhyd.2022.107755
[19]	张意锋, 邓云, 赵艳云. 双循环高静压对鱿鱼原肌球蛋白二、三级结构的影响[J]. 中国食品学报,2018,18(5):45−50. [ZHANG Y F, DENG Y, ZHAO Y Y. Effect of two-cycle high hydrostatic pressure on the secondary and tertiary structures of squid Tropomyosin Tod p1[J]. Journal of Chinese Institute of Food Science and Technology,2018,18(5):45−50.
[20]	OGUNWOLU S O, HENSHAW F O, MOCK H P, et al. Functional properties of protein concentrates and isolates produced from cashew (Anacardium occidentale L.) nut[J]. Food Chemistry,2009,115(3):852−858. doi: 10.1016/j.foodchem.2009.01.011
[21]	SHEN P, GAO Z, XU M, et al. The impact of hempseed dehulling on chemical composition, structure properties and aromatic profile of hemp protein isolate[J]. Food Hydrocolloids,2020,106:105889. doi: 10.1016/j.foodhyd.2020.105889
[22]	MALOMO S A, HE R, ALUKO R E. Structural and functional properties of hemp seed protein products[J]. Journal of Food Science,2014,79(8):C1512−C1521. doi: 10.1111/1750-3841.12537
[23]	FEVZIOGLU M, OZTURK O K, HAMAKER B R, et al. Quantitative approach to study secondary structure of proteins by FT-IR spectroscopy, using a model wheat gluten system[J]. International Journal of Biological Macromolecules,2020,164:2753−2760. doi: 10.1016/j.ijbiomac.2020.07.299
[24]	张永金, 胡艳红, 葛武鹏, 等. 母乳、牛乳与主要小品种乳蛋白质组成及乳清蛋白二级结构比较[J]. 食品安全质量检测学报,2022,13(15):4779−4786. [ZHANG Y J, HU Y H, GE W P, et al. Comparative study on the composition of protein and secondary structure of whey protein in human milk, milk and main small varieties of milk[J]. Journal of Food Safety & Quality,2022,13(15):4779−4786.
[25]	DENG Y, HUANG L, ZHANG C, et al. Physicochemical and functional properties of Chinese quince seed protein isolate[J]. Food Chemistry,2019,283:539−548. doi: 10.1016/j.foodchem.2019.01.083
[26]	YU M, ZENG M, QIN F, et al. Physicochemical and functional properties of protein extracts from Torreya grandis seeds[J]. Food Chemistry,2017,227:453−460. doi: 10.1016/j.foodchem.2017.01.114
[27]	TAN M, XU J, GAO H, et al. Effects of combined high hydrostatic pressure and pH-shifting pretreatment on the structure and emulsifying properties of soy protein isolates[J]. Journal of Food Engineering,2021,306:110622. doi: 10.1016/j.jfoodeng.2021.110622
[28]	BAKWO BASSOGOG C B, NYOBE C E, NGUI S P, et al. Effect of heat treatment on the structure, functional properties and composition of Moringa oleifera seed proteins[J]. Food Chemistry,2022,384:132546. doi: 10.1016/j.foodchem.2022.132546
[29]	WANG Q, XIONG Y L. Processing, nutrition, and functionality of hempseed protein: A review[J]. Comprehensive Reviews in Food Science and Food Safety,2019,18(4):936−952. doi: 10.1111/1541-4337.12450
[30]	SHEVKANI K, SINGH N, KAUR A, et al. Structural and functional characterization of kidney bean and field pea protein isolates: A comparative study[J]. Food Hydrocolloids,2015,43:679−689. doi: 10.1016/j.foodhyd.2014.07.024
[31]	YIN S W, TANG C H, CAO J S, et al. Effects of limited enzymatic hydrolysis with trypsin on the functional properties of hemp (Cannabis sativa L.) protein isolate[J]. Food Chemistry,2007,106(3):1004−1013.
[32]	AMAGLIANI L, O'REGAN J, KELLY A L, et al. The composition, extraction, functionality and applications of rice proteins: A review[J]. Trends in Food Science & Technology,2017,64:1−12.