均质联合酶解对大豆蛋白纳米颗粒结构与功能特性的影响

王金鸽; 蔡勇建; 刘俊梅; 赵强忠

doi:10.13386/j.issn1002-0306.2022090179

均质联合酶解对大豆蛋白纳米颗粒结构与功能特性的影响

doi: 10.13386/j.issn1002-0306.2022090179

王金鸽^1,,
蔡勇建²,
刘俊梅^1, ,,
赵强忠^2, ,

1.
吉林农业大学食品科学与工程学院，吉林长春 130118
2.
华南理工大学食品科学与工程学院，广东广州 510640

基金项目: 国家自然科学基金（32072148，32101866）；“十四五”国家重点研发计划（2021YFD2100102）。

详细信息

作者简介:
王金鸽（1997−），女，硕士研究生，研究方向：食品乳浊体系，E-mail：jgwang97@163.com

通讯作者:
刘俊梅（1973−），女，博士，教授，研究方向：功能性食品，E-mail：spring430817@163.com

赵强忠（1976−），男，博士，教授，研究方向：食品乳浊体系，E-mail：qzzhao@scut.edu.cn

中图分类号: TS201.7
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- 被引次数: 0
出版历程
- 收稿日期: 2022-09-19
- 网络出版日期: 2023-05-21
- 刊出日期: 2023-07-01

Effect of Homogenization Assisted with Enzymatic Treatment on the Structural and Functional Properties of Soybean Protein Nanoparticles

1.
School of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China
2.
School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China

摘要

摘要: 本文以商业大豆分离蛋白（Soy protein isolate，SPI）为原料，分别通过酶解、均质联合酶解制备了蛋白纳米颗粒（Soy protein nanoparticles，SPNPs），对比分析了SPNPs的粒径、多相分散系数及微观形态、傅里叶红外光谱、内源荧光等结构特征，以及内部作用力、表面疏水性、Zeta电位、两亲特性、乳化性与起泡性等物化特性。研究发现：SPI粒径较大（230.00 nm），低水解度（3%）酶解和均质联合酶解处理制备的SPNPs粒径减小（64.20~144.80 nm），呈小球形。二级结构分析表明均质联合酶解制备SPNPs的α-螺旋/β-折叠比例（约45%）较高。与单一酶解所制SPNPs相比，均质联合酶解制备的SPNPs在中性条件时具有更强负电荷（−33 mV），表面疏水性更高，乳化和起泡性能更强。内部作用力结果表明疏水相互作用主导了纳米颗粒结构的形成，氢键和二硫键分别为维持纳米颗粒外部和内部结构的主要作用力。上述结果表明均质协同酶解处理为绿色制备多功能蛋白纳米颗粒提供了新的解决思路。
- 大豆蛋白纳米颗粒（SPNPs） /
- 均质-酶解 /
- 结构 /
- 功能特性
Abstract: The commercial soy protein isolate (SPI) was used as raw material to prepare soy protein nanoparticles (SPNPs) through either single enzymatic treatment or homogenization assisted with enzymatic treatment. The structural characteristics of particle size, polydispersity index, morphology, Fourier transform infrared and endogenous fluorescence spectra of SPNPs, as well as the physicochemical properties of the pattern of intra-particle interactive forces, surface hydrophobicity, Zeta-potential, amphipathy, emulsifying, and foaming properties of SPNPs were comparably analyzed. It was found that the SPI with large particle size (230.00 nm), the SPNPs prepared by either enzymatic or homogenization assisted with enzymatic treatment at low degree (3%) of hydrolysis (DH) were spherical and showed smaller size distributions with z-average size from 64.20 to 144.80 nm. The analysis of secondary structure implied that SPNPs prepared by homogenization assisted with enzymatic treatment showed an increased ratio of α-helix/β-sheet to a narrow range of around 45%. Compared with SPNPs prepared by single enzymatic treatment, the SPNPs showed a stronger negative charge (−33 mV) under neutral conditions and higher H₀, suggesting better emulsifying and foaming properties of the present SPNPs. Meanwhile, the results of the interactive force indicated that the hydrophobic interactions mainly dominated the structure formation of SPNPs, along with hydrogen bonds and disulfide bonds mainly stabilizing the external and internal structure of nanoparticles, respectively. The above results indicated that homogenization assisted with enzymatic treatment provided new solutions for the green preparation of multifunctional protein nanoparticles.
- soy protein nanoparticles (SPNPs) /
- homogenization assisted with enzymatic treatment /
- structural /
- functional properties

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图 1 不同酶酶解SPI的水解度随时间的变化规律

Figure 1. The DH of SPI by different enzymes as a function of hydrolysis time

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图 2 SPI及SPNPs的FE-SEM形貌观察

Figure 2. The FE-SEM microscopic observation of SPI and SPNPs

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图 3 SPI及SPNPs的内源荧光光谱图

Figure 3. The endogenous fluorescence spectra of SPI and SPNPs

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图 4 大豆蛋白纳米颗粒在不同蛋白变性溶剂中的外观和粒径

Figure 4. The appearance and particle size of SPNPs in different protein denaturant solutions

注：1：去离子水；2：DTT；3：SDS；4：尿素；5：DTT+SDS；6：DTT+尿素；7：SDS+尿素；8：DTT+SDS+尿素；A、B、C、D分别代表：SPNP-_E3、SPNP-_E6、SPNP-_HE3、SPNP-_HE6；不同小写字母代表同一样品在不同溶剂中的粒径差异显著（P<0.05）。

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图 5 SPI及SPNPs的表面疏水性

Figure 5. The H₀ of SPI and SPNPs

注：不同字母代表样品间差异显著（P<0.05）；图6同。

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图 6 SPI及SPNPs的Zeta电位

Figure 6. The Zeta potential of SPI and SPNPs

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表 1 SPI及SPNPs的平均粒径和多相分散系数

Table 1. The z-average size and PDI of SPI and SPNPs

样品	平均粒径（nm）	PDI
SPI	230.00±1.54^b	0.78±0.03^a
SPIH	181.70±21.50^c	0.47±0.01^b
SPNP-_E3	144.80±1.41^d	0.27±0.00^c
SPNP-_E6	260.30±8.44^a	0.29±0.01^c
SPNP-_HE3	64.20±4.32^f	0.20±0.00^c
SPNP-_HE6	106.10±2.36^e	0.24±0.00^c
注：同列不同小写字母代表样品间差异显著（P<0.05）；表2~表4同。

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表 2 SPI及SPNPs的二级结构组成

Table 2. The composition of secondary structures of SPI and SPNPs

样品	α-螺旋（%）	β-折叠（%）	β-转角（%）	无规则卷曲（%）	α-螺旋/β-折叠（%）
SPI	14.77±0.05^d	43.34±0.01^cd	20.44±0.30^b	21.46±0.01^a	34.08±0.39^e
SPIH	15.70±0.16^c	43.75±0.22^c	22.99±0.60^a	17.56±0.23^c	35.89±0.33^d
SPNP-_E3	17.11±0.10^b	43.40±0.03^cd	20.33±0.20^b	19.14±0.34^b	39.42±0.11^c
SPNP-_E6	16.71±0.02^b	46.35±0.50^a	18.64±0.03^cd	18.30±0.76^bc	36.05±0.31^d
SPNP-_HE3	20.09±0.05^a	42.59±0.23^d	19.05±0.06^c	18.27±0.12^bc	47.17±0.19^a
SPNP-_HE6	19.55±0.04^a	44.78±0.10^b	18.11±0.17^d	17.56±0.02^c	43.66±0.21^b

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表 3 SPI及SPNPs的溶解性及持水/持油性

Table 3. The solubility and WHC/OHC of SPI and SPNPs

样品	溶解性（%）	持水性（g/g）	持油性（g/g）
SPI	44.36±0.01^c	13.19±1.04^a	2.19±0.49^c
SPIH	68.20±1.02^b	9.79±0.35^b	4.48±0.77^b
SPNP-_E3	95.06±1.11^a	接近0	4.68±0.15^b
SPNP-_E6	93.75±0.56^a	接近0	4.57±0.12^b
SPNP-_HE3	95.50±0.06^a	接近0	6.83±0.67^a
SPNP-_HE6	93.70±0.92^a	接近0	6.40±0.42^a

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表 4 SPI及SPNPs的乳化性、乳液稳定性和起泡性、泡沫稳定性

Table 4. The EAI, ESI and FAI, FSI of SPI and SPNPs

样品	乳化性（m²/g）	乳液稳定性（min）	起泡性（%）	泡沫稳定性（%）
SPI	39.66±0.43^f	18.44±0.14^e	54.44±1.57^d	69.42±2.00^b
SPIH	46.57±0.51^e	33.37±0.66^d	67.78±0.77^c	86.88±0.30^a
SPNP-_E3	58.73±0.57^c	51.17±0.49^c	70.78±0.46^c	60.75±4.14^c
SPNP-_E6	51.23±0.81^d	29.63±0.56^de	80.47±1.31^b	81.66±1.69^a
SPNP-_HE3	66.48±0.50^a	126.27±3.36^a	100.09±3.49^a	85.89±0.05^a
SPNP-_HE6	61.64±0.67^b	71.80±0.13^b	108.28±2.13^a	90.56±0.79^a

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