添加山茶油对肌原纤维蛋白凝胶特性的影响

doi:10.3864/j.issn.0578-1752.2021.20.017

摘要/Abstract

摘要：

【目的】研究山茶油添加对凝胶中肌原纤维蛋白（myofibrillar protein,MP）特性的影响以及结构与凝胶特性间的关系,为其在鸡肉糜类制品中的应用提供理论依据。【方法】取鸡胸肉于4℃下提取MP,在添加山茶油条件下测定其流变特性,水浴加热制成凝胶后测定其硬度、保水性和水分布特性的变化;通过扫描电镜、激光拉曼光谱、表面疏水性（S₀-ANS）、总巯基含量、差示扫描量热法（DSC）等分析,探讨山茶油对MP凝胶微观结构、二级和三级结构以及热变性的影响,并分析山茶油添加量、凝胶特性和MP结构之间的相关性,揭示山茶油添加促使凝胶特性改变的原因。【结果】MP凝胶的硬度、保水性和束缚水的百分比（PT₂₁）随着山茶油添加量的增加而显著提高（P<0.05）,在4%时分别达到最大值116.60 g、95.77%和80.05%,5%时有所下降。山茶油添加量为4%时,MP加热过程中的储能模量（G'）最高,凝胶弹性最大,此时扫描电镜下的MP混合凝胶网络结构均匀、致密,表面粗糙度最低。山茶油的添加（0–5%）影响MP的二级结构,其α-螺旋含量从39.94%显著降低至26.07%（P<0.05）,β-折叠含量从22.04%显著升高至27.40%（P<0.05）,β-转角和无规则卷曲含量呈上升趋势。山茶油添加量增加到4%时,MP凝胶的表面疏水性达到最高水平,总巯基含量最低。结果表明山茶油的添加促进MP分子内部疏水性氨基酸残基暴露和分子间二硫键的生成,三级结构发生变化,更高的β-折叠含量和疏水相互作用增强MP分子间的聚集和交联。相关性分析表明,山茶油添加量与MP凝胶特性及蛋白质构象的变化显著相关（P<0.05）。DSC结果表明添加的山茶油能与MP相互作用,降低MP的变性热焓。【结论】添加山茶油对MP凝胶特性和结构有显著影响。添加山茶油改变MP的微环境并与MP发生作用,使MP更容易变性并导致其二级结构展开以及三级结构改变,进而改善MP凝胶特性。当山茶油添加量为4%时,混合凝胶特性和三维网状结构最佳。

关键词: 山茶油, 肌原纤维蛋白, 凝胶特性, 结构

Abstract:

【Objective】The effects of camellia oil addition on the properties and the relationship between the properties and structure of myofibrillar protein (MP) gel were studied. The purpose of the research was to provide a theoretical basis for the application of camellia oil in chicken products. 【Method】The MP was extracted from chicken breast at 4℃. The MP heat-induced gels in different camellia oil addition were prepared. The changes of hardness, water holding capacity (WHC), moisture distribution of MP gel and rheological properties of MP were measured. Furthermore, the effects of camellia oil on the microstructure, secondary structure were determined by scanning electron microscopy (SEM) and Raman spectroscopy, respectively. Surface hydrophobicity and total sulfhydryl content were used to study the tertiary structure of MP in gels. Meanwhile, the differential scanning calorimetry (DSC) was applied to characterize the thermal properties of MP. Finally, the correlation between camellia oil addition, gel properties and MP structures were analyzed. 【Result】After 4% camellia oil addition, the hardness, WHC and the area fraction of bound water (PT₂₁) of MP gel increased and reached to maximum values of 116.60 g, 95.77% and 80.05%, respectively. However, those values decreased as camellia oil addition increased to 5%. For camellia oil content of 4%, the storage modulus (G') and elasticity of MP were the highest, while the surface roughness was the lowest. Meanwhile, MP gels showed more uniform, and compact network structure compared with other groups. It was also found that secondary structure of MP was significantly affected by camellia oil addition (0-5%). The α-helix content decreased from 39.94% to 26.07%, and the β-sheet content increased from 22.04% to 27.40% with increased camellia oil addition (P<0.05). The content of β-turn and random coil showed increasing trends. The S₀-ANS of MP reached the highest value, and the total sulfhydryl content was the lowest after 4% camellia oil addition. The results showed that the addition of camellia oil promoted the exposure of hydrophobic amino acid residues in protein molecules and the formation of disulfide bond, and changed the tertiary structure of MP in gels. The enhanced aggregation and cross-linking between MP molecules were due to the higher β-sheet content and hydrophobic interaction. The correlation analysis indicated that camellia oil addition was significantly correlated with the changes in the gel properties and protein structure (P<0.05). The results of DSC showed that camellia oil could interact with MP and reduce the denaturation enthalpy of MP. 【Conclusion】The camellia oil addition had a significant effect on the gel properties and structures of MP. The addition of camellia oil changed the microenvironment of MP and interacted with MP, which made the degeneration of MP easier. The secondary structure of MP was unfolded and the tertiary structure was affected after camellia oil addition, which improved the properties of MP gel. When camellia oil concentration was 4%, the properties and three-dimensional network structure of MP composite gel were the best.

Key words: camellia oil, myofibrillar protein, gel properties, structure

韩柯颖,冯潇,杨玉玲,李珊珊,魏苏萌,陈玉敏. 添加山茶油对肌原纤维蛋白凝胶特性的影响[J]. 中国农业科学, 2021, 54(20): 4446-4455.

HAN KeYing,FENG Xiao,YANG YuLing,LI ShanShan,WEI SuMeng,CHEN YuMin. Effects of Camellia Oil on the Properties of Myofibrillar Protein Gel[J]. Scientia Agricultura Sinica, 2021, 54(20): 4446-4455.

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山茶油含量 Camellia oil content (%)	α-螺旋 α-helix (%)	β-折叠 β-sheet (%)	β-转角 β-turn (%)	无规则卷曲 Random coil (%)
0	39.94±0.91a	22.04±0.32d	19.77±0.40e	18.25±0.63b
1	36.52±1.19b	23.13±1.00c	20.41±0.42e	19.94±2.53ab
2	33.85±0.74c	24.55±0.34b	21.93±0.61d	19.67±0.45ab
3	30.49±0.92d	25.05±0.28b	23.06±0.52c	21.39±1.06a
4	28.35±0.46e	26.63±0.95a	23.96±0.35b	21.06±0.59a
5	26.07±0.76f	27.40±0.08a	25.22±0.40a	21.31±0.70a

	山茶油添加量 Camellia oil content	硬度 Hardness	保水性 WHC	PT₂₁ PT₂₁	储能模量(65℃) G'(65℃)	α-螺旋 α-helix	β-折叠 β-sheet	表面疏水性 S₀-ANS	总巯基 Total SH
山茶油添加量Camellia oil content	1
硬度 Hardness	0.889*	1
保水性 WHC	0.846*	0.919**	1
PT₂₁	0.437	0.711	0.833*	1
G' (65℃)	0.804	0.971**	0.939**	0.819*	1
α-螺旋α-helix	-0.997**	-0.896*	-0.865*	-0.482	-0.815*	1
β-折叠β-sheet	0.994**	0.886*	0.871*	0.463	0.810	-0.989**	1
表面疏水性 S₀-ANS	0.644	0.848*	0.933**	0.956**	0.935**	-0.675	0.664	1
总巯基 Total SH	-0.928**	-0.973**	-0.966**	-0.727	-0.960**	0.939**	-0.932**	-0.879*	1

山茶油含量 Camellia oil content (%)	T_d1			T_d2
山茶油含量 Camellia oil content (%)	T_Onset(℃)	T_End(℃)	△H (J∙g^-1)	T_Onset(℃)	T_End(℃)	△H (J∙g^-1)
0	55.14±0.54a	62.89±0.22a	0.183±0.007a	70.50±0.35b	71.89±1.05a	0.023±0.003a
1	54.73±0.43a	62.69±0.11ab	0.158±0.013b	70.97±0.06a	72.19±0.25a	0.024±0.004a
2	54.64±0.03a	62.31±0.12b	0.152±0.004b	70.97±0.06a	72.24±0.10a	0.022±0.003a
3	54.54±0.03a	62.34±0.42ab	0.155±0.017b	70.81±0.02ab	72.08±0.32a	0.021±0.001a
4	54.45±0.04a	62.30±0.42b	0.154±0.006b	70.77±0.21ab	71.90±0.27a	0.023±0.003a
5	54.39±0.10a	62.37±0.28ab	0.143±0.011b	68.41±0.12c	71.42±0.55a	0.023±0.001a