不同添加顺序对β-乳球蛋白与EGCG及葡萄糖三元复合物的结构和功能的影响

摘要/Abstract

摘要： 摘要：采用碱法和美拉德反应将β-乳球蛋白（β-LG）、表没食子儿茶素没食子酸酯（EGCG）及葡萄糖（Glc）通过不同添加顺序获得两种共价复合物为β-LG-EGCG-Glc con和β-LG-Glc-EGCG con，与直接混合形成的非共价复合物β-LG-EGCG-Glc mix和β-LG-Glc-EGCG mix进行对比研究，探讨不同添加顺序对三元复合物的结构和功能性质的影响。研究结果发现不同添加顺序对共价复合物的影响大于非共价复合物。β-LG-EGCG-Glc con与β-LG-Glc-EGCG con在结构和功能性质上有较大差异，而β-LG-EGCG-Glc mix和β-LG-Glc-EGCG mix在结构和功能上差别不大。SDS-PAGE电泳和紫外吸收光谱结果显示不同添加顺序对三元复合物的共价结合过程具有较大影响。结构表征发现，共价复合物的荧光猝灭和疏水性都高于非共价复合物。相较于β-LG-EGCG-Glc con，β-LG-Glc-EGCG con有更强的猝灭程度以及更高的疏水性，而β-LG-EGCG-Glc mix与β-LG-Glc-EGCG mix两者差距不明显。此外，两种共价复合物的功能性质都优于非共价复合物。β-LG-Glc-EGCG con的乳化性及乳化稳定性、起泡性以及起泡稳定性都优于β-LG-EGCG-Glc con，而非共价结合物之间差距不大。

关键词: 关键词：β-乳球蛋白, 三元复合物, 功能性质, 结构, 共价结合, 非共价结合

Abstract: Abstract: The β-lactoglobulin (β-LG), epigallocatechin gallate (EGCG) and glucose (Glc) were conjugated via different addition sequences under the alkali method and the Maillard reaction to obtain two covalent complexes, named as β-LG-EGCG-Glc con and β-LG-Glc-EGCG con. Compared with β-LG-EGCG-Glc mix and β-LG-Glc-EGCG mix. The effects of different addition sequences on the structural and functional properties of ternary complexes was investigated. The results showed that the different addition order had a greater effect on the covalent complex than the non-covalent complex. The structure and function of β-LG-EGCG-Glc con and β-LG-Glc-EGCG con were significant difference, while there was little difference between β-LG-EGCG-Glc mix and β-LG-Glc-EGCG mix. SDS-PAGE electrophoresis and UV absorption spectroscopy results displayed that the different order of addition had a great influence on the covalent bonding process of ternary complexes. Structural characterization revealed that the fluorescence quenching and surface hydrophobicity of the covalent complexes were higher than the non-covalent complexes. Compared with β-LG-EGCG-Glc con, β-LG-Glc-EGCG con had a stronger degree of fluorescence quenching and higher surface hydrophobicity, while there was not obvious changes between β-LG-EGCG-Glc mix and β-LG-Glc-EGCG mix. In addition, the functional properties of the two covalent complexes were superior to those of the non-covalent complexes. The emulsification and emulsion stability, foaming and foaming stability of β-LG-Glc-EGCG con were superior to those of β-LG-EGCG-Glc con, but little difference between the two direct mixing complexes.

Key words: Key words: β-lactoglobulin, ternary complex, functional properties, structure, covalent bonding, non-covalent binding

中图分类号:

TS201.2

姚文俊周磊付珊琳钟俊桢刘成梅. 不同添加顺序对β-乳球蛋白与EGCG及葡萄糖三元复合物的结构和功能的影响[J]. 食品科学, 0, (): 0-0.

参考文献

[1] Hattori M, Numamoto K, Kobayashi K, et al.Functional changes in β-lactoglobulin by conjugation with cationic saccharides[J]. Journal of Agricultural and Food Chemistry, 200048(6):2050-2056. DOI: 10.1021/jf991105s.[J].Journal of Agricultural and Food Chemistry, 2000, 48(6):2050-2056 [2]Bin Y.Qin,Maria CBewley,Creamer L K,et al. Structural Basis of the Tanford Transition of Bovine β-Lactoglobulin ?,?[J].Biochemistry, 1998, 37(40):14014-14023 [3]Liang L, Subirade M.Beta-lactoglobulinfolic acid complexes: formation,characterization,and biological implication[J].Journal of Physical Chemistry B, 2010, 114(19):6707-6712 [4]Livney Y D.Milk proteins as vehicles for bioactives[J].Current Opinion in Colloid & Interface Science, 2010, 15(1):73-83 [5]Li M, Hagerman A E.Role of the flavan-3-ol and galloyl moieties in the interaction of (-)-epigallocatechin gallate with serum albumin[J].Journal of Agricultural and Food Chemistry, 2014, 62(17):3768-3775 [6]Li M, Ma Y, Ngadi M O.Binding of curcumin to β-lactoglobulin and its effect on antioxidant characteristics of curcumin[J].Food Chemistry, 2013, 141(2):1504-1511 [7]Bourvellec C L, Renard C M G C.Interactions between Polyphenols and Macromolecules: Quantification Methods and Mechanisms[J].Critical Reviews in Food Science and Nutrition, 2012, 52(3):213-248 [8]Ozdal T, Capanoglu E, Altay F.A review on protein–phenolic interactions and associated changes[J].Food Research International, 2013, 51(2):954-970 [9]Le B C, Renard C M.Interactions between polyphenols and macromolecules: quantification methods and mechanisms[J].Critical Reviews in Food Science and Nutrition, 2012, 52(3):213-248 [10] Liu F, Ma C, Gao Y, et al.Food-Grade Covalent Complexes and Their Application as Nutraceutical Delivery Systems: A Review[J]. Comprehensive Reviews in Food Science and Food Safety, 201616: 76-95. DOI: 10.1111/1541-4337.12229.[J].Comprehensive Reviews in Food Science and Food Safety, 2016, 16(1):76-95 [11]Yi J, Zhang Y, Liang R, et al.Beta-carotene chemical stability in nanoemulsions was improved by stabilized with beta-lactoglobulin-catechin conjugates through free radical method[J].Journal of Agricultural and Food Chemistry, 2015, 63(1):297-303 [12] Jia Z, Zheng M, Tao F, et al.Effect of covalent modification by (?)-epigallocatechin-3-gallate on physicochemical and functional properties of whey protein isolate[J].LWT-Food Science and Technology, 2016, 66:305-310. DOI: 10.1016/j.lwt.2015.10.054.[J].LWT-FOOD SCIENCE AND TECHNOLOGY, 2016, 66:305-310 [13]Xia S, Li Y, Zhao Q, et al.Probing Conformational Change of Bovine Serum Albumin–Dextran Conjugates under Controlled Dry Heating[J].Journal of Agricultural and Food Chemistry, 2015, 63(16):4080-4086 [14]Harbertson J F, Yuan C, Mireles M S, et al.Glucose,fructose and sucrose increase the solubility of protein-tannin complexes and at high concentration,glucose and sucrose interfere with bisulphite bleaching of wine pigments[J].Food Chemistry, 2013, 138(1):556-563 [15]Soares S, Mateus N, Freitas V D.Carbohydrates Inhibit Salivary Proteins Precipitation by Condensed Tannins[J].Journal of Agricultural and Food Chemistry, 2012, 60(15):3966-3972 [16]Aberkane L, Jasniewski J, Gaiani C, et al.Structuration mechanism of β-lactoglobulin– acacia gum assemblies in presence of quercetin[J].Food Hydrocolloids, 2012, 29(1):9-20 [17]Yang W, Xu C, Liu F, et al.Fabrication mechanism and structural characteristics of the ternary aggregates by lactoferrin,pectin,and (-)-epigallocatechin gallate using multispectroscopic methods[J].Journal of Agricultural and Food Chemistry, 2015, 63(20):5046-5054 [18]Rawel H M, Czajka D, Rohn S, et al.Interactions of different phenolic acids and flavonoids with soy proteins[J].International Journal of Biological Macromolecules, 2002, 30(3):137-150 [19]Liu F, Sun C, Wang D, et al.Glycosylation improves the functional characteristics of chlorogenic acid-lactoferrin conjugates[J].Rsc Advances, 2015, 5(95):78215-78228 [20]Laemmli U K.Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4[J].Nature, 1970, 227(5259):680-685 [21]Ajandouz E H, Tschiape L S, Dalle O F, et al.Effects of pH on caramelization and Maillard reaction kinetics in fructose-lysine model systems[J].Journal of Food Science, 2010, 66(7):926-931 [22]Liu C M, Zhong J Z, Liu W, et al.Relationship between functional properties and aggregation changes of whey protein induced by high pressure microfluidization[J].Journal of Food Science, 2011, 76(4):E341-E347 [23]Pearce K N, Kinsella J E.Emulsifying properties of proteins: Evaluation of a turbidimetric technique[J].Journal of Agricultural and Food Chemistry, 1978, 26(3):716-723 [24]彭倩, 颜小燕, 李雨婷, 等.盐提和碱提酸沉两种方法提取腰果蛋白的结构及其功能性质分析[J].食品科学, 2017, 38(23):75-81 [25]Kroll J, Harshadrai M R, Rohn S.Reactions of Plant Phenolics with Food Proteins and Enzymes under Special Consideration of Covalent Bonds[J].Food science and Technology Research, 2003, 9(3):205-218 [26]Spotti M J, Perduca M J, Piagentini A, et al.Gel mechanical properties of milk whey protein–dextran conjugates obtained by Maillard reaction[J].Food Hydrocolloids, 2013, 31(1):26-32 [27]Ali M, Homann T, Kreisel J, et al.Characterization and modeling of the interactions between coffee storage proteins and phenolic compounds[J].Journal of Agricultural and Food Chemistry, 2012, 60(46):11601-11608 [28] 张捷.乳蛋白和三种活性成分间相互作用及对活性成分的保护作用[D]. 江南大学, 2014. [29]Vigo M S, Malec L S, Gomez R G, et al.Spectrophotometric assay using o -phthaldialdehyde for determination of reactive lysine in dairy products[J].Food Chemistry, 1992, 44(5):363-365 [30]Liu Y, Zhao G, Zhao M, et al.Improvement of functional properties of peanut protein isolate by conjugation with dextran through Maillard reaction[J].Food Chemistry, 2012, 131(3):901-906 [31]And A P, Lichan E C Y.Comparison of Protein Surface Hydrophobicity Measured at Various pH Values Using Three Different Fluorescent Probes[J].Journal of Agricultural and Food Chemistry, 2000, 48(2):328-334 [32]Collini M, D' Alfonso L, Baldini G.New insight on β-lactoglobulin binding sites by 1-anilinonaphthalene-8-sulfonate fluorescence decay[J].Protein Science, 2000, 9(10):1968-1974 [33]Collini M, D' Alfonso L, Molinari H, et al.Competitive binding of fatty acids and the fluorescent probe 1-8-anilinonaphthalene sulfonate to bovine beta-lactoglobulin[J].Protein Science, 2003, 12(8):1596-1603 [34]Kontopidis G, Holt C, Sawyer L.Invited review: beta-lactoglobulin: binding properties,structure,and function[J].Journal of Dairy Science, 2004, 87(4):785-796 [35]Aydemir L Y, G?kbulut A A, Baran Y, et al.Bioactive,functional and edible film-forming properties of isolated hazelnut (Corylus avellana L) meal proteins[J].Food Hydrocolloids, 2014, 36(2014):130-142 [36] 陈豪, 钟俊桢, 黄宗兰, 等.三种方法提取的澳洲坚果蛋白的功能性质与构象的关系 [J]食品科学2018-01-12. 食品科学