氧化对肌原纤维蛋白流变学特性及结构的影响
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摘要
以鸡肉肌原纤维蛋白(myofibrillar protein,MP)作为研究对象,利用脂肪氧化酶-亚油酸体系研究了蛋白质氧化对MP加热过程中的流变性质、凝胶状态下的二级结构和疏水相互作用的影响,并探讨了加热过程中的流变性质、凝胶状态下蛋白质的二级结构与疏水相互作用的内在联系。研究表明,与其他亚油酸浓度处理的样品相比,亚油酸浓度为2 mmol/L的MP样品的G′值(>68℃)最大。随着亚油酸的加入,α-螺旋含量从未氧化时的48.31%一直降低到亚油酸10 mmol/L时的最低值29.57%,而β-折叠含量则从14.75%缓慢升高到22.14%,β-转角含量也随着氧化程度的升高,从16.28%增加到21.88%,无规则卷曲的含量总体呈现升高的趋势。随着氧化程度的升高,MP凝胶的疏水相互作用先增加后降低,在2 mmol/L达到最大值,同时G′值(>68℃)在2 mmol/L时达到最大,这表明疏水相互作用的变化与MP凝胶中蛋白质二级结构的变化具有密切的相关性。
The study was designed to reveal the effect of protein oxidation on rheological properties during heating,hydrophobic interaction and secondary structure of chicken myofibrillar protein (MP) gel using lipoxygenase-linoleic acid system. The internal relationship between MP rheological properties during heating,protein secondary structure in gel state and hydrophobic interaction was revealed. The results showed that,MP samples with a linoleic acid concentration of 2 mmol/L had the largest G′ value (>68 ℃) compared with other linoleic acid concentrations. With the addition of linoleic acid,the α-helix content decreased from 48.31% in the unoxidized state to the lowest value of 29.57% in the case of linoleic acid 10 mmol/L,while the β-sheet content gradually increased from 14.75% to 22.14%. The β-turn content also increased from 16.28% to 21.88% with the increase of oxidation degree,and the content of random coil generally showed an increasing trend. As the degree of oxidation increased,the hydrophobic interaction of the MP gel firstly increased and then decreased,reaching a maximum at 2 mmol/L,while the G′ value (>68 ℃) reached a maximum at 2 mmol/L,which showed that the change of hydrophobic interaction was closely related to the change of protein secondary structure in MP gel.
The study was designed to reveal the effect of protein oxidation on rheological properties during heating,hydrophobic interaction and secondary structure of chicken myofibrillar protein (MP) gel using lipoxygenase-linoleic acid system. The internal relationship between MP rheological properties during heating,protein secondary structure in gel state and hydrophobic interaction was revealed. The results showed that,MP samples with a linoleic acid concentration of 2 mmol/L had the largest G′ value (>68 ℃) compared with other linoleic acid concentrations. With the addition of linoleic acid,the α-helix content decreased from 48.31% in the unoxidized state to the lowest value of 29.57% in the case of linoleic acid 10 mmol/L,while the β-sheet content gradually increased from 14.75% to 22.14%. The β-turn content also increased from 16.28% to 21.88% with the increase of oxidation degree,and the content of random coil generally showed an increasing trend. As the degree of oxidation increased,the hydrophobic interaction of the MP gel firstly increased and then decreased,reaching a maximum at 2 mmol/L,while the G′ value (>68 ℃) reached a maximum at 2 mmol/L,which showed that the change of hydrophobic interaction was closely related to the change of protein secondary structure in MP gel.
引文
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[2]管斌,林宏.食品蛋白质化学 [M].北京:化工出版社,2005:140-162.
[3]Estevez M,Ventanas S,Cava R.Protein oxidation in frankfurters with increasing levels of added rosemary essential oil:Effect on color and texture deterioration[J].Journal of Food Science,2010,70(7):427-432.
[4]Lund M N,Lametsch R,Hviid M S,et al.High-oxygen packaging atmosphere influences protein oxidation and tenderness of porcine longissimus dorsi during chill storage[J].Meat Science,2007,77(3):295-303.
[5]Liu G,Xiong Y L,Butterfield D A.Chemical,physical,and gel-forming properties of oxidized myofibrils and whey-and soy-protein isolates[J].Journal of Food Science,2010,65(5):811-818.
[6]Xiong Y L,Lou X,Wang C,et al.Protein extraction from chicken myofibrils irrigated with various polyphosphate and NaCl solutions [J].Journal of Food Science,2000,65(1):96-100.
[7]杨玉玲,游远,彭晓蓓,等.加热对鸡胸肉肌原纤维蛋白结构与凝胶特性的影响[J].中国农业科学,2014,47(10):2013-2020.
[8]Gornall A G,Bardawill C J,David M M.Determination of serum proteins by means of the biuret reaction[J].Journal of Biological Chemistry,1949,177(2):751-766.
[9]Uchida K,Kanematsu M,Morimitsu Y,et al.Acrolein is aproduct of lipid peroxidation reaction.Formation of free acrolein and its conjugate with lysine residues in oxidized low density lipoproteins[J].The Journal of Biological Chemistry,1998,273(26):16058-16066.
[10]Li-Chan E C Y.The applications of Raman spectroscopy in food science[J].Trends in Food Science and Technology,1996,7(11):361-370.
[11]Li-Chan E,Nakai S,Hirotsuka M.Raman spectroscopy as a probe of protein structure in food systems[J].Protein Structure-Function Relationships in Foods,1994:163-197.
[12]Liu G,Xiong Y L.Electrophoretic pattern,thermal denaturation,and in vitro digestibility of oxidized myosin[J].Journal of Agricultural and Food Chemistry,2000,48(3):624-630.
[13]Li-Chan E C Y.The applications of Raman spectroscopy in food science[J].Trends in Food Science and Technology,1996,7(11):361-370.
[14]Nonaka M,Li-Chan E,Nakai S.Raman spectroscopic study of thermally induced gelation of whey proteins[J].Journal of Agricultural and Food Chemistry,1993,41(8):1176-1181.
[15]Alix A J P,Pedanou G,Berjot M.Fast determination of the quantitative secondary structure of proteins by using some parameters of Raman amide I band[J].Journal of Molecular Structure,1988,174(174):159-164.
[16]吴伟.蛋白质氧化对大豆蛋白结构和凝胶特性的影响[D].无锡:江南大学,2010.
[17]Tamamizukato S,Wong J Y,Jairam V,et al.Modification by acrolein,a component of tobacco smoke and age-related oxidative stress,mediates functional impairment of human apolipoprotein E[J].Biochemistry,2007,46(28):8392-8400.
[18]Li S J,King A J.Lipid oxidation and myosin denaturation in dark chicken meat[J].Journal of Agricultural and Food Chemistry,1996,44(10):3080-3084.
[19]Wu W,Wu X,Hua Y.Structural modification of soy protein by the lipid peroxidation product acrolein[J].LWT-Food Science and Technology,2010,43(1):133-140.
[20]Gao J,Bosco D A,Powers E T,et al.Localized thermodynamic coupling between hydrogen bonding and microenvironment polarity substantially stabilizes proteins[J].Nature Structural & Molecular Biology,2009,16(7):684-690.
[21]陈琼.鸡胸肉肌原纤维蛋白与脂肪替代品混合凝胶形成作用力研究[D].南京:南京财经大学,2012.