豌豆蛋白对牛肉盐溶蛋白理化性质及二级结构的影响
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摘要
以牛背最长肌为原料,研究豌豆蛋白对其盐溶蛋白十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate and polyacrylamide gelelectrophoresis, SDS-PAGE)、溶解度、表面疏水性、巯基、二硫键含量、乳化性等理化性质及二级结构的影响,并探讨了豌豆蛋白与盐溶蛋白凝胶形成过程中化学作用力类型。结果表明,SDS-PAGE电泳中,豌豆蛋白中的vicilin(7S)、legA和legB分别与盐溶蛋白中的肌动蛋白、肌钙蛋白T和肌钙蛋白C结合。当豌豆蛋白添加质量浓度为0.08 g/mL时,盐溶蛋白的表面疏水性上升明显(P<0.05);二硫键含量在豌豆蛋白添加浓度超过0.12 g/mL后,迅速上升(P<0.05)。随豌豆蛋白添加浓度的增加,盐溶蛋白总巯基和表面活性巯基含量下降,浊度和乳化性上升;溶解度呈先升高后下降趋势,在添加浓度为0.04 g/mL时最大。豌豆蛋白添加质量浓度与α-螺旋、表面活性巯基含量等均呈显著负相关,与β-折叠、β-转角、无规则卷曲、二硫键含量、表面疏水性等呈显著正相关;疏水相互作用是豌豆蛋白-牛肉盐溶蛋白凝胶网络结构形成中最主要化学作用力。
Salt-soluble proteins were extracted from bovine longissimus thoracis muscles(BLTM). Effects of pea protein on physicochemical properties(SDS-PAGE, solubility, surface hydrophobicity, sulfhydryl group, disulfide bond content, emulsifying properties) and secondary structures of salt-soluble proteins from BLTM were studied. Additionally, the types of chemical bonds formed during the gelling of pea protein and salt-soluble proteins were explored. The SDS-PAGE results indicated that vicilin(7 S), legA, and legB in pea protein were bound to actin, troponin T, and troponin C in salt-soluble proteins. When 0.08 g/mL pea protein was added, the surface hydrophobicity of salt-soluble protein increased significantly(P<0.05). Moreover, the disulfide bond content increased rapidly when more than 0.12 g/mL pea protein was added(P<0.05). As the concentration of pea protein increasing, the contents of both total sulfhydryl and surface active sulfhydryl of salt-soluble proteins decreased, while their turbidity and emulsibility increased. Furthermore, the solubility of salt-soluble proteins increased first and then decreased, with the maximal reached when 0.04 g/mL pea protein was added. Additionally, the amount of pea protein added was negatively correlated with α-helix and surface active sulfhydryl content, and it was positively correlated with β-folding, β-rotation, irregular curl, disulfide bond content, and surface hydrophobicity of salt soluble proteins. Besides, hydrophobic interaction was the most important chemical force when forming pea protein-BLTM salt soluble proteins gel network structure.
Salt-soluble proteins were extracted from bovine longissimus thoracis muscles(BLTM). Effects of pea protein on physicochemical properties(SDS-PAGE, solubility, surface hydrophobicity, sulfhydryl group, disulfide bond content, emulsifying properties) and secondary structures of salt-soluble proteins from BLTM were studied. Additionally, the types of chemical bonds formed during the gelling of pea protein and salt-soluble proteins were explored. The SDS-PAGE results indicated that vicilin(7 S), legA, and legB in pea protein were bound to actin, troponin T, and troponin C in salt-soluble proteins. When 0.08 g/mL pea protein was added, the surface hydrophobicity of salt-soluble protein increased significantly(P<0.05). Moreover, the disulfide bond content increased rapidly when more than 0.12 g/mL pea protein was added(P<0.05). As the concentration of pea protein increasing, the contents of both total sulfhydryl and surface active sulfhydryl of salt-soluble proteins decreased, while their turbidity and emulsibility increased. Furthermore, the solubility of salt-soluble proteins increased first and then decreased, with the maximal reached when 0.04 g/mL pea protein was added. Additionally, the amount of pea protein added was negatively correlated with α-helix and surface active sulfhydryl content, and it was positively correlated with β-folding, β-rotation, irregular curl, disulfide bond content, and surface hydrophobicity of salt soluble proteins. Besides, hydrophobic interaction was the most important chemical force when forming pea protein-BLTM salt soluble proteins gel network structure.
引文
[1] 周纷,谷大海,徐家慧,等.淀粉对鸡胸肉盐溶性蛋白乳化特性的影响[J].食品科学,2016,37(15):7-12.
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[3] 周向军,朱敏涛,袁毅君.赤藓糖醇对豌豆分离蛋白结构和功能特性的影响[J].食品工业科技,2018,39(8):73-77;84.
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[5] 何希强,肖怀秋,李玉珍.豌豆蛋白酶法水解及产物特性研究[J].粮油食品科技,2012,20(5):8-11.
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[7] 马宁,魏姜勉.豌豆蛋白的改性及其开发利用研究进展[J].中国市场,2015,32:231-233.
[8] 冯婷,孙京新,吴振,等.花生浓缩蛋白对鸡胸肉盐溶蛋白热诱导凝胶性能的影响[J].现代食品科技,2015,31(10):97-102;128.
[9] 张巍.大豆分离蛋白(SPI)与猪肉盐溶蛋白共凝胶的研究[D].哈尔滨:哈尔滨商业大学,2011.
[10] 李先保,程千赫.大豆分离蛋白对兔肉盐溶蛋白凝胶特性的影响[J].食品与发酵工业,2010,36(5):133-139.
[11] 白一凡,王辉,杨震,等.豌豆蛋白加工特性及在乳化香肠中的应用[J].肉类研究,2014,28(12):14-16.
[12] 杨震,曲超,贡慧,等.豌豆蛋白组织化挤压工艺参数优化及其在肉制品中的应用[J].延边大学农学学报,2016,38(4):317-324.
[13] SU Y K,BOWERS J A,ZAYAS J F.Physical characteristics and microstructure of reduced-fat frankfurters as affected by salt and emulsified fats stabilized with nonmeat proteins [J].Journal of Food Science,2000,65(1):123-128.
[14] CHEN H,XU S Y,WANG Z.Interaction between flaxseed gum and meat protein [J].Journal of Food Engineering,2007,80(4):1 051-1 059
[15] 孔鹏,王志耕.3种淀粉对鸡肉糜盐溶蛋白特性影响及其配方研究[J].肉类研究,2011,25(6):9-15.
[16] 栗俊广,蒋爱民,白艳红,等.木薯淀粉对猪肉盐溶蛋白凝胶特性的影响[J].现代食品科技,2016,32(11):123-127,55.
[17] 郭尧君.蛋白质电泳实验技术[M].北京:科学出版社,2005:123-157.
[18] LAEMMLI U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227:680-685.
[19] 尚永彪,夏杨毅,张彩霞,等.磷酸盐对PSE猪肉肌原纤维蛋白溶胶及凝胶性质的影响[J].食品科学,2010,31(1):38-42.
[20] 魏朝贵,吴菊清,邵俊花,等.KCl和MgCl2部分替代NaCl对猪肉肌原纤维蛋白乳化凝胶特性的影响[J].食品科学,2014,35(5):89-95.
[21] CHELH I,GATELLIER P,SANTELHOUTELLIER V.Technical note:a simplified procedure for myofibril hydrophobicity determination [J].Meat Science,2006,74(4):681-683.
[22] 邵俊花.猪肉蛋白质构象变化与保油保水性关系研究[D].南京:南京农业大学,2012.
[23] THANNHAUSER T W,KONISHI Y,SCHERAGA H A,et al.Analysis for disulfide bonds in peptides and proteins [J].Methods in Enzymology,1987,143(1):115-119.
[24] 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.
[25] GóMEZ-GUILLéN M C,BORDEíAS A J,MONTERO P.Chemical interactions of nonmuscle proteins in the network of sardine (sardine pilchardus) muscle gels [J].LWT-Food Science and Technology,1997,30(6):602-608.
[26] YANG H,ZHANG W,LI T,et al.Effect of protein structure on water and fat distribution during meat gelling [J].Food Chemistry,2016,204:239-245.
[27] 刘英丽,谢良需,丁立,等.小麦麸膳食纤维对猪肉肌原纤维蛋白凝胶功能特性的影响[J].食品科学,2016,37(19):15-23.
[28] GATEHOUSE J A,LYCETT G W,CROY R R,et al.The post-translational proteolysis of the subunits of vicilin from pea (Pisum sativum L.)[J].Biochemical Journal,1982,207(3):629-632.
[29] BARAC M,CABRILO S,PESIC M,et al.Profile and functional properties of seed proteins from six pea (Pisum sativum) genotypes [J].International Journal of Molecular Sciences,2010,11(12):4 973-4 990.
[30] O’KANE F E,HAPPE R P,VEREIJKEN J M,et al.Characterization of pea vicilin 2:Consequences of compositional heterogeneity on heat-induced gelation behavior[J].Journal of Agricultural and Food Chemistry,2004,52(10):3 149-3 154.
[31] CHAN J K,GILL T A.Thermal aggregation of mixed fish myosins [J].Journal of Agriculture and Food Chemistry,1994,42(12):2 649-2 655.
[32] 闫海鹏.不同种类肉肌原纤维蛋白功能特性的研究[D].南京:南京农业大学,2013.
[33] 宋江宁,李炜疆.蛋白质二硫键的分布特征[J].无锡轻工大学学报,2002,21(5):464-467.
[34] 张兴,杨玉玲,马云,等.pH对肌原纤维蛋白及其热诱导凝胶非共价键作用力与结构的影响[J].中国农业科学,2017,50(3):564-573.
[35] CHOI S M,MA C Y.Structural characterization of globulin from common buckwheat (Fagopyrum esculentum Moench) using circular dichroism and Raman spectroscopy [J].Food Chemistry,2007,102(1):150-160.
[36] 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):2 013-2 020.
[37] WU Z,BERTRAM H C,BCKER U,et al.Myowater dynamics and protein secondary structural changes as affected by heating rate in three pork qualities:a combined FT-IR microspectroscopic and 1H NMR relaxometry study [J].Journal of Agricultural and Food Chemistry,2007,55:3 990-3 997.
[38] WANG S F,SMITH D M.Dynamic rheological properties and secondary structure of chicken breast myosin as influenced by isothermal heating [J].Journal of Agricultural and Food Chemistry,1994,42(7):1 434-1 439.
[39] 康怀彬,邹良亮,张慧芸,等.高温处理对牛肉蛋白质化学作用力及肌原纤维蛋白结构的影响[J/OL].食品科学:http://kns.cnki.net/kcms/detail/11.2206.TS.20180622.1127.164.html
[2] 彭增起.肌肉盐溶蛋白质溶解性和凝胶特性研究[D].南京:南京农业大学,2005.
[3] 周向军,朱敏涛,袁毅君.赤藓糖醇对豌豆分离蛋白结构和功能特性的影响[J].食品工业科技,2018,39(8):73-77;84.
[4] SWIATECKA D,NARBAD A,RIDGWAY K P,et al.The study on the impact of glycated pea proteins on human intestinal bacteria [J].International Journal of Food Microbiology,2011,145(1):267-272.
[5] 何希强,肖怀秋,李玉珍.豌豆蛋白酶法水解及产物特性研究[J].粮油食品科技,2012,20(5):8-11.
[6] COSTA G E D A,QUEIROZ-MONICI K D S,REIS S M P M,et al.Chemical composition,dietary fibre and resistant starch contents of raw and cooked pea,common bean,chickpea and lentil legumes[J].Food Chemistry,2006,94(3):327-330.
[7] 马宁,魏姜勉.豌豆蛋白的改性及其开发利用研究进展[J].中国市场,2015,32:231-233.
[8] 冯婷,孙京新,吴振,等.花生浓缩蛋白对鸡胸肉盐溶蛋白热诱导凝胶性能的影响[J].现代食品科技,2015,31(10):97-102;128.
[9] 张巍.大豆分离蛋白(SPI)与猪肉盐溶蛋白共凝胶的研究[D].哈尔滨:哈尔滨商业大学,2011.
[10] 李先保,程千赫.大豆分离蛋白对兔肉盐溶蛋白凝胶特性的影响[J].食品与发酵工业,2010,36(5):133-139.
[11] 白一凡,王辉,杨震,等.豌豆蛋白加工特性及在乳化香肠中的应用[J].肉类研究,2014,28(12):14-16.
[12] 杨震,曲超,贡慧,等.豌豆蛋白组织化挤压工艺参数优化及其在肉制品中的应用[J].延边大学农学学报,2016,38(4):317-324.
[13] SU Y K,BOWERS J A,ZAYAS J F.Physical characteristics and microstructure of reduced-fat frankfurters as affected by salt and emulsified fats stabilized with nonmeat proteins [J].Journal of Food Science,2000,65(1):123-128.
[14] CHEN H,XU S Y,WANG Z.Interaction between flaxseed gum and meat protein [J].Journal of Food Engineering,2007,80(4):1 051-1 059
[15] 孔鹏,王志耕.3种淀粉对鸡肉糜盐溶蛋白特性影响及其配方研究[J].肉类研究,2011,25(6):9-15.
[16] 栗俊广,蒋爱民,白艳红,等.木薯淀粉对猪肉盐溶蛋白凝胶特性的影响[J].现代食品科技,2016,32(11):123-127,55.
[17] 郭尧君.蛋白质电泳实验技术[M].北京:科学出版社,2005:123-157.
[18] LAEMMLI U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227:680-685.
[19] 尚永彪,夏杨毅,张彩霞,等.磷酸盐对PSE猪肉肌原纤维蛋白溶胶及凝胶性质的影响[J].食品科学,2010,31(1):38-42.
[20] 魏朝贵,吴菊清,邵俊花,等.KCl和MgCl2部分替代NaCl对猪肉肌原纤维蛋白乳化凝胶特性的影响[J].食品科学,2014,35(5):89-95.
[21] CHELH I,GATELLIER P,SANTELHOUTELLIER V.Technical note:a simplified procedure for myofibril hydrophobicity determination [J].Meat Science,2006,74(4):681-683.
[22] 邵俊花.猪肉蛋白质构象变化与保油保水性关系研究[D].南京:南京农业大学,2012.
[23] THANNHAUSER T W,KONISHI Y,SCHERAGA H A,et al.Analysis for disulfide bonds in peptides and proteins [J].Methods in Enzymology,1987,143(1):115-119.
[24] 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.
[25] GóMEZ-GUILLéN M C,BORDEíAS A J,MONTERO P.Chemical interactions of nonmuscle proteins in the network of sardine (sardine pilchardus) muscle gels [J].LWT-Food Science and Technology,1997,30(6):602-608.
[26] YANG H,ZHANG W,LI T,et al.Effect of protein structure on water and fat distribution during meat gelling [J].Food Chemistry,2016,204:239-245.
[27] 刘英丽,谢良需,丁立,等.小麦麸膳食纤维对猪肉肌原纤维蛋白凝胶功能特性的影响[J].食品科学,2016,37(19):15-23.
[28] GATEHOUSE J A,LYCETT G W,CROY R R,et al.The post-translational proteolysis of the subunits of vicilin from pea (Pisum sativum L.)[J].Biochemical Journal,1982,207(3):629-632.
[29] BARAC M,CABRILO S,PESIC M,et al.Profile and functional properties of seed proteins from six pea (Pisum sativum) genotypes [J].International Journal of Molecular Sciences,2010,11(12):4 973-4 990.
[30] O’KANE F E,HAPPE R P,VEREIJKEN J M,et al.Characterization of pea vicilin 2:Consequences of compositional heterogeneity on heat-induced gelation behavior[J].Journal of Agricultural and Food Chemistry,2004,52(10):3 149-3 154.
[31] CHAN J K,GILL T A.Thermal aggregation of mixed fish myosins [J].Journal of Agriculture and Food Chemistry,1994,42(12):2 649-2 655.
[32] 闫海鹏.不同种类肉肌原纤维蛋白功能特性的研究[D].南京:南京农业大学,2013.
[33] 宋江宁,李炜疆.蛋白质二硫键的分布特征[J].无锡轻工大学学报,2002,21(5):464-467.
[34] 张兴,杨玉玲,马云,等.pH对肌原纤维蛋白及其热诱导凝胶非共价键作用力与结构的影响[J].中国农业科学,2017,50(3):564-573.
[35] CHOI S M,MA C Y.Structural characterization of globulin from common buckwheat (Fagopyrum esculentum Moench) using circular dichroism and Raman spectroscopy [J].Food Chemistry,2007,102(1):150-160.
[36] 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):2 013-2 020.
[37] WU Z,BERTRAM H C,BCKER U,et al.Myowater dynamics and protein secondary structural changes as affected by heating rate in three pork qualities:a combined FT-IR microspectroscopic and 1H NMR relaxometry study [J].Journal of Agricultural and Food Chemistry,2007,55:3 990-3 997.
[38] WANG S F,SMITH D M.Dynamic rheological properties and secondary structure of chicken breast myosin as influenced by isothermal heating [J].Journal of Agricultural and Food Chemistry,1994,42(7):1 434-1 439.
[39] 康怀彬,邹良亮,张慧芸,等.高温处理对牛肉蛋白质化学作用力及肌原纤维蛋白结构的影响[J/OL].食品科学:http://kns.cnki.net/kcms/detail/11.2206.TS.20180622.1127.164.html
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