复合保护剂与牛IgG的相互作用机制研究

刘妍妍; 陈文璐; 李建立; 李凤; 孙正瀚; 栾晶华

doi:10.13386/j.issn1002-0306.2022050111

复合保护剂与牛IgG的相互作用机制研究

doi: 10.13386/j.issn1002-0306.2022050111

1.
黑龙江八一农垦大学食品学院，黑龙江大庆 163319
2.
山东得益乳业股份有限公司技术中心，山东淄博 265200
3.
威海威职国有资产运营有限公司，山东威海 264200

基金项目: 功能性液态奶的研制及产业化（2019ZX07B02）。

详细信息

作者简介:
刘妍妍（1979−），女，硕士，副教授，研究方向：畜产品精深加工，E-mail：spxylyy@126.com

通讯作者:
刘妍妍（1979−），女，硕士，副教授，研究方向：畜产品精深加工，E-mail：spxylyy@126.com

中图分类号: TS252.5
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- 文章访问数: 133
- HTML全文浏览量: 26
- PDF下载量: 9
- 被引次数: 0
出版历程
- 收稿日期: 2022-05-11
- 网络出版日期: 2023-04-05
- 刊出日期: 2023-06-01

Study on the Interaction Mechanism between Compound Protective Agent and Bovine IgG

1.
College of Food Science, Heilongjiang Bayi Agricultural University, Daqing 163319, China
2.
Technology Center, Shandong Deyi Dairy Products Co., Ltd., Zibo 265200, China
3.
Weihai Vocational College State-owned Property Operation Co., Ltd., Weihai 264200, China

摘要

摘要: 本研究综合利用紫外吸收光谱法、荧光光谱法、傅里叶变换红外光谱法、圆二色谱法、拉曼光谱法、扫描电镜法和差示扫描量热法研究复合保护剂与IgG的相互作用机制以及IgG结构的变化。结果表明复合保护剂对IgG的肽骨架影响较小，而对氨基酸残基的微环境影响较大，使IgG氨基酸残基所处的微环境极性增强；复合保护剂与IgG的相互作用主要涉及氢键、静电作用力和疏水作用力，并引起IgG的二级结构从α-螺旋结构向β-折叠、β-转角和无规卷曲结构转变，使IgG的二级结构趋于无序化，IgG二硫键构型由g-g-g构型逐渐转变为g-g-t构型和t-g-t构型。扫描电镜表明，复合保护剂在IgG分子表面形成不规则的棱形晶体复合物；差示扫描量热法表明，复合保护剂使IgG的变性温度从71.76 ℃升高到87.22 ℃，IgG的热焓变从6.51 J/g降低到4.68 J/g。说明复合保护剂使IgG分子的构象发生转变，从而增强了IgG的热稳定性。
- IgG /
- 复合保护剂 /
- 作用机制 /
- 光谱法
Abstract: In this study, ultraviolet absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, circular dichroism, Raman spectroscopy, scanning electron microscopy and differential scanning calorimetry were used to study the interaction of composite protectants with IgG mechanisms and changes in IgG structure. The results showed that the compound protective agent had little effect on the peptide skeleton of IgG, but had a great effect on the microenvironment of amino acid residues, which enhanced the polarity of the microenvironment of amino acid residues in IgG. The interaction of compound protective agent and IgG mainly involved hydrogen bond, electrostatic force and hydrophobic force, and caused the secondary structure from to changed from α-helix structure to β-folding, β-turning and random curl structure of IgG tended to disorder the secondary structure. The IgG disulfide bond configurations gradually changed from g-g-g to g-g-t and t-g-t. The results of SEM showed that the compound protective agent formed irregular prismatic crystal complexes on the surface of IgG molecules, differential scanning calorimetry showed that the compound protective agent increased the denaturation temperature of IgG from 71.76 ℃ to 87.22 ℃, and decreased the enthalpy change of IgG from 6.51 J/g to 4.68 J/g. The results showed that the conformation of IgG molecule was changed by compound protective agent, which enhanced the thermal stability of IgG.
- IgG /
- compound protective agent /
- mechanism /
- spectroscopy

HTML全文

图 1 IgG和复合保护剂-IgG作用体系的同步荧光光谱图

Figure 1. Synchronous fluorescence spectrogram of IgG and compound protective agent-IgG interaction system

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图 2 IgG和复合保护剂-IgG作用体系的紫外光谱图

Figure 2. Ultraviolet second derivative spectrogram of IgG and compound protective agent-IgG interaction system

注：A：紫外光谱图；B：紫外二阶导数光谱图：a为tyr的λ_max（～λ₂₈₈）和λ_min（～λ₂₈₄）纵坐标间距；b为trp的λ_max（～λ₂₉₅）和λ_min（～λ₂₉₁）纵坐标间距。

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图 3 IgG和复合保护剂-IgG作用体系的三维荧光光谱等高线图

Figure 3. Three-dimensional fluorescence contour map of IgG and compound protective agent-IgG interaction system

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图 4 IgG和复合保护剂-IgG作用体系的红外光谱图

Figure 4. FTIR spectrogram of IgG and compound protective agent-IgG interaction system

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图 5 IgG和复合保护剂-IgG作用体系的圆二光谱图

Figure 5. Circular dichroism spectrogram of IgG and compound protective agent-IgG interaction system

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图 6 IgG和复合保护剂-IgG作用体系的拉曼光谱图

Figure 6. Raman spectrogram of IgG and compound protective agenton-IgG interaction system

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图 7 IgG和复合保护剂-IgG作用体系的扫描电镜图

Figure 7. SEM spectrogram of IgG and compound protective agent-IgG interaction system

注：图中a~c和d~f分别代表IgG和复合保护剂-IgG，放大倍数分别为300×、3000×、8000×。

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表 1 IgG含量和活性指标的变化

Table 1. Changes in IgG content and activity indicators

指标	IgG	复合保护剂-IgG
IgG含量（µg/mL）	590±150^b	1830±130^a
IgG活性（IU/L）	74.62±1.12^b	463.84±1.23^a
注：同行不同字母表示差异显著（P<0.05）。

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表 2 IgG和复合保护剂-IgG作用体系的荧光激发-发射矩阵光谱

Table 2. Fluorescent EEM spectral characteristics of IgG and compound protective agent-IgG interaction system

样品	Peak 1		Peak 2
样品	λ_ex/λ_em（nm/nm）	荧光强度F	λ_ex/λ_em（nm/nm）	荧光强度F
IgG	280/340	563.87	235/340	308.45
复合保护剂-IgG	280/344	433.74	235/341	235.53

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表 3 IgG和复合保护剂-IgG作用体系二级结构的相对含量

Table 3. Secondary structure composition of IgG and compound protective agent-IgG interaction system

样品	α-螺旋（%）	β-折叠（%）	β-转角（%）	无规卷曲（%）
IgG	25.10±0.06^a	21.70±0.04^b	18.40±0.03^b	39.40±0.05^b
复合保护剂-IgG	19.50±0.07^b	26.70±0.06^a	19.90±0.05^a	44.70±0.07^a
注：同列不同小写字母表示显著性差异（P<0.05）；表5~表7同。

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表 4 IgG和复合保护剂-IgG作用体系的拉曼特征峰位及峰位归属

Table 4. Assignment of characteristic bands in the Raman spectra of IgG and compound protective agent-IgG interaction system

波数（cm⁻¹）	峰位归属
500～500	s-s伸缩振动
760	色氨酸
850/830	酪氨酸
1003	苯丙氨酸
1220～1330	酰胺III带
1600～1700	酰胺I带

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表 5 IgG和复合保护剂-IgG作用体系的酪氨酸、色氨酸微环境

Table 5. Tyrosine and tryptophan microenvironment of IgG and compound protective agent-IgG interaction system

样品	I_{850 cm}⁻¹/I_{830 cm}⁻¹	I_{760 cm}⁻¹
IgG	1.21±0.06^a	0.90±0.08^a
复合保护剂-IgG	1.09±0.04^b	0.91±0.05^a

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表 6 IgG和复合保护剂-IgG作用体系中二硫键构型组成含量

Table 6. Configurational compositions of disulfide bonds in IgG and compound protective agent-IgG interaction system

样品	g-g-g构型（%）	g-g-t构型（%）	t-g-t构型（%）
IgG	64.50±0.11^a	10.90±0.06^b	24.60±0.08^b
复合保护剂-IgG	26.40±0.05^b	28.10±0.07^a	45.50±0.09^a

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表 7 IgG和复合保护剂-IgG作用体系的DSC分析

Table 7. DSC analysis of IgG and compound protective agent-IgG interaction system

样品	T_d（℃）	ΔH（J/g）
IgG	71.76±0.08^b	6.51±0.05^a
复合保护剂-IgG	87.22±0.06^a	4.68±0.07^b

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