微波加热对肌球蛋白理化特性的影响
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摘要
试验研究比较了肌球蛋白在不同微波加热速率条件下的理化性质的变化,测定其浊度、溶解度、巯基含量、表面疏水性和二硫键等指标,并通过SDS-PAGE分析,探讨微波加热条件下肌球蛋白的变性及聚集过程。结果表明:当微波功率密度为8 W/mL和2.67 W/mL时,溶液浊度及肌球蛋白溶解度变化趋势及对应的温度范围差异明显。在8 W/mL下,肌球蛋白分子在短时间(80 s)内边变性边聚集,二硫键形成量较少且时间滞后;而在2.67 W/mL下,肌球蛋白分子则呈现逐渐变性而后聚集的趋势,与常规蛋白质凝胶形成过程类似。推测可能是快速升温及电磁波高频作用导致蛋白质结构变化、变性及聚集行为发生变化。
The changes in physicochemical properties of myosin at two microwave power levels were studied. The denaturation and aggregation of myosin during microwave heating were researched through determination of turbidity of myosin solution, solubility of myosin, sulfhydryl content, surface hydrophobicity and disulfide bond in myosin, and analysis of SDS-PAGE electrophoresis. The results showed when microwave power densities were 8 W/mL and 2.67 W/mL, respectively,the changing trends of solution turbidity of myosin solution and solubility of myosin during microwave heating and the temperature ranges exhibited obvious difference. Denaturation and aggregation of myosin were carried out simultaneously in80 s, and less content of disulfide bond was produced and delayed in time at the microwave power densities of 8 W/mL. At 2.67 W/m L, myosin aggregated after it denatured gradually, which was similar as traditional gelation procedure of protein. It was surmised that denaturation and aggregation behaviors of myosin were changed as a result of change in protein structure, which was caused by rapid heating and energy of high frequency microwave.
The changes in physicochemical properties of myosin at two microwave power levels were studied. The denaturation and aggregation of myosin during microwave heating were researched through determination of turbidity of myosin solution, solubility of myosin, sulfhydryl content, surface hydrophobicity and disulfide bond in myosin, and analysis of SDS-PAGE electrophoresis. The results showed when microwave power densities were 8 W/mL and 2.67 W/mL, respectively,the changing trends of solution turbidity of myosin solution and solubility of myosin during microwave heating and the temperature ranges exhibited obvious difference. Denaturation and aggregation of myosin were carried out simultaneously in80 s, and less content of disulfide bond was produced and delayed in time at the microwave power densities of 8 W/mL. At 2.67 W/m L, myosin aggregated after it denatured gradually, which was similar as traditional gelation procedure of protein. It was surmised that denaturation and aggregation behaviors of myosin were changed as a result of change in protein structure, which was caused by rapid heating and energy of high frequency microwave.
引文
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[13] LIU W, LANIER T C. Rapid(microwave)heating rate effects on texture, fat/water holding, and microstructure of cooked comminuted meat batters[J]. Food Research International,2016(81):108-113.
[14]潘锦锋,沈慧星,尤娟,等.草鱼肌原纤维蛋白加热过程中理化特性的变化[J].中国农业大学学报, 2009, 14(6):17-22.
[15] ANMING W, MEIZHANG W, QIUYAN W, et al. Stable and efficient immobilization technique of aldolase under consecutive microwave irradiation at low temperature[J].Bioresource Technology, 2011, 102(2):469-474.
[16] LIU W. Studies on heat-induced gelation of myofibrillar protein pastes with emphasis on stabilization of added fat[J].Dissertations&Theses-Gradworks, 2012.
[17]潘锦锋,沈慧星,尤娟,等.草鱼肌原纤维蛋白加热过程中理化特性的变化[J].中国农业大学学报, 2009, 14(6):17-22.
[18] LEFEVRE F, FAUCONNEAU B, AND J W T, et al.Thermal denaturation and aggregation properties of atlantic salmon myofibrils and myosin from white and red muscles[J].Journal of Agricultural&Food Chemistry, 2007, 55(12):4761.
[19]白云,谌启亮,周光宏,等.温度对兔腰大肌肌球蛋白质凝胶特性的影响[J].江苏农业学报, 2011, 27(2):405-409.
[20] SANDOVAL W N, PHAM V C, LILL J R. Recent developments in microwave-assisted protein chemistries-can this be integrated into the drug discovery and validation process?[J]. Drug Discovery Today, 2008, 13(23/24):1075-1081.
[2]董秋颖,杨玉玲,许婷.从质构学角度研究肌原纤维蛋白凝胶形成的作用力[J].食品与发酵工业, 2009, 35(5):45-49.
[3]付湘晋.白鲢鱼脱腥及其低盐鱼糜制备的研究[D].无锡:江南大学, 2009.
[4] WANG S F, SMITH D M. Heat-induced denaturation and rheological properties of chicken breast myosin and F-Actin in the presence and absence of pyrophosphate[J]. Journal of Agricultural&Food Chemistry, 1994, 42(12):2665-2670.
[5]徐幸莲.兔骨骼肌肌球蛋白热诱导凝胶特性及成胶机制研究[D].南京:南京农业大学, 2003.
[6] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J]. Nature, 1970,227(5259):680-685.
[7]郭尧君.蛋白质电泳实验技术[M].北京:科学出版社,2005.
[8]尹寿伟.芸豆蛋白的物化修饰及相关构效机理研究[D].广州:华南理工大学, 2009.
[9]韩敏义.兔骨骼肌肌球蛋白浊度、溶解度及热诱导凝胶强度研究[D].南京:南京农业大学, 2003.
[10] CHELH I, GATELLIER P, SANTé-LHOUTELLIER V.Technical note:A simplified procedure for myofibril hydrophobicity determination[J]. Meat Science, 2006, 74(4):681-683.
[11]曾宪明,徐幸莲,白云,等.兔肌球蛋白热诱导凝胶过程中的物理化学变化[J].食品科学, 2008, 29(4):149-153.
[12] THANNHAUSER T W, KONISHI Y, SCHERAGA H A.Analysis for disulfide bonds in peptides and proteins[J]. Methods in Enzymology, 1987, 143(1):115.
[13] LIU W, LANIER T C. Rapid(microwave)heating rate effects on texture, fat/water holding, and microstructure of cooked comminuted meat batters[J]. Food Research International,2016(81):108-113.
[14]潘锦锋,沈慧星,尤娟,等.草鱼肌原纤维蛋白加热过程中理化特性的变化[J].中国农业大学学报, 2009, 14(6):17-22.
[15] ANMING W, MEIZHANG W, QIUYAN W, et al. Stable and efficient immobilization technique of aldolase under consecutive microwave irradiation at low temperature[J].Bioresource Technology, 2011, 102(2):469-474.
[16] LIU W. Studies on heat-induced gelation of myofibrillar protein pastes with emphasis on stabilization of added fat[J].Dissertations&Theses-Gradworks, 2012.
[17]潘锦锋,沈慧星,尤娟,等.草鱼肌原纤维蛋白加热过程中理化特性的变化[J].中国农业大学学报, 2009, 14(6):17-22.
[18] LEFEVRE F, FAUCONNEAU B, AND J W T, et al.Thermal denaturation and aggregation properties of atlantic salmon myofibrils and myosin from white and red muscles[J].Journal of Agricultural&Food Chemistry, 2007, 55(12):4761.
[19]白云,谌启亮,周光宏,等.温度对兔腰大肌肌球蛋白质凝胶特性的影响[J].江苏农业学报, 2011, 27(2):405-409.
[20] SANDOVAL W N, PHAM V C, LILL J R. Recent developments in microwave-assisted protein chemistries-can this be integrated into the drug discovery and validation process?[J]. Drug Discovery Today, 2008, 13(23/24):1075-1081.