食品与发酵工业 >

2020 , Vol. 46 >Issue 11: 61 - 68

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.022733

研究报告

牛血清蛋白与邻苯二酚、间苯三酚相互作用的机理探究

  • 褚千千 ,
  • 艾健 ,
  • 方佳琪 ,
  • 韩秋煜 ,
  • 包斌 ,
  • 孙永军
展开
  • 1(上海海洋大学 食品学院,上海,201306)
    2(食品科学与工程国家级实验教学示范中心(上海海洋大学),上海,201306)
    3(好当家集团有限公司,山东 荣成,264305)
硕士研究生(包斌副教授和孙永军高级工程师为共同通讯作者, E-mail:bbao@shou.com;Sun29889@163.com)

收稿日期: 2019-11-06

  网络出版日期: 2020-06-24

收起

Study on the mechanism of interactions between bovine serum albuminwith o-dihydroxybenzene and m-trihydroxybenzene

  • CHU Qianqian ,
  • AI Jian ,
  • FANG Jiaqi ,
  • HAN Qiuyu ,
  • BAO Bin ,
  • SUN Yongjun
Expand
  • 1(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China)
    2(National Experimental Teaching Demonstration Center for Food Science and Engineering (Shanghai Ocean University),Shanghai 201306,China)
    3(Good Master Group Co., Ltd., Rongcheng 264305, China)

Received date: 2019-11-06

  Online published: 2020-06-24

Fold

摘要

为探究酚类化合物与蛋白质的相互作用机理,分别用氧化的邻苯二酚 (oxidized o-dihydroxybenzene, OOP)、间苯三酚 (oxidized m-trihydroxybenzene, OMP)和未氧化的邻苯二酚(OP)、间苯三酚(m-trihydroxybenzene, MP)和牛血清蛋白(bovine serum protein, BSA)在常温下反应。分析了BSA与OP、MP、OOP、OMP反应前后游离氨基、巯基、非共价相互作用力、表面疏水性的变化;应用傅里叶变换红外光谱 (Fourier transform infrared spectroscopy, FTIR)、圆二色谱 (circular dichroism, CD)、差式扫描量热仪 (differential scanning calorimeter, DSC) 表征构象的变化。结果显示:BSA与OP、MP 相互作用后,游离氨基、巯基含量的变化不显著,但是OOP、OMP可使BSA游离氨基、巯基含量显著降低; OP、MP可与BSA形成氢键,但离子键没有显著变化; OP与BSA反应后,疏水相互作用显著升高,而MP导致疏水相互作用显著降低; OP、MP、OOP、OMP都会使BSA表面疏水性显著降低; FTIR和CD结果显示, OP、MP、OOP、OMP都会导致BSA二级结构的变化,表现为α-螺旋含量降低,β-折叠、β-转角、无规卷曲含量上升;DSC结果表明, OP、MP、OOP、OMP的引入提高了BSA的热变性温度。综上,邻苯二酚、间苯三酚经过氧化后与BSA发生共价相互作用,而未氧化的邻苯二酚、间苯三酚可与BSA发生非共价相互作用,这些相互作用可能改变蛋白质的功能性质,从而影响蛋白质在食品中的应用。

关键词： 邻苯二酚; 间苯三酚; 蛋白质; 共价相互作用; 非共价相互作用

本文引用格式

褚千千 , 艾健 , 方佳琪 , 韩秋煜 , 包斌 , 孙永军 . 牛血清蛋白与邻苯二酚、间苯三酚相互作用的机理探究[J]. 食品与发酵工业, 2020 , 46(11) : 61 -68 . DOI: 10.13995/j.cnki.11-1802/ts.022733

Abstract

This study aimed to investigate the interaction mechanism between phenolic compounds and proteins. The o-dihydroxybenzene (OP) and m-trihydroxybenzene (MP) were oxidized under alkaline conditions in the presence of oxygen to obtain oxidized o-dihydroxybenzene (OOP) and oxidized m-trihydroxybenzene (OMP), which were respectively reacted with oxidized and unoxidized OP, MP and BSA at room temperature. The changes of free amino, sulfhydryl, non-covalent interaction and surface hydrophobicity between BSA and OP, MP, OOP and OMP were analyzed. Changes were characterized by Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD) and differential scanning calorimeter (DSC). The results showed that the content of free amino and sulfhydryl groups did not change significantly between BSA and OP and MP, but OOP and OMP could significantly reduce the content of free amino and sulfhydryl groups in BSA.AndthereductionofOOP-BSA was greater than that of OMP-BSA. Moreover, MP can form hydrogen bonds with BSA, but the ionic bond did not change significantly and the increase of MP-BSA hydrogen bond was greater than that of OP-BSA. After the reaction between OP and BSA, the hydrophobic interaction increased significantly. While MP caused the hydrophobic interaction to decrease significantly. Furthermore, OP, MP, OOP, OMP could significantly reduce the hydrophobicity of BSA surface and the reduction of OP-BSA, OOP-BSA was greater than MP-BSA, OMP-BSA. FTIR and CD results showed that OP, MP, OOP, OMP could change the secondary structure of BSA which was characterized by the decreased α-helix content and increased β-sheet, β-turn and random coil. The effects of OP and OOP were greater than those of MP and OMP. DSC results showed that the introduction of OP, MP, OOP and OMP increased the thermal denaturation temperature of BSA. These results indicated that catechol and phloroglucinol undergo oxidative interaction with BSA and unoxidized catechol and phloroglucinol could interact non-covalently with bovine serum albumin. These interactions may alter the functional properties of the protein, thereby affecting the application of the protein in food.

Key words： o-dihydroxybenzene; m-trihydroxybenzene; protein; covalent interaction; non-covalent interaction

参考文献

[1] ZIEGLER V, FERREIRA C D, HOFFMANN J F, et al. Effects of moisture and temperature during grain storage on the functional properties and isoflavone profile of soy protein concentrate[J]. Food Chemistry, 2018, 242: 37-44.
[2] DE CASTRO R J S, DOMINGUES M A F, OHARA A, et al. Whey protein as a key component in food systems: Physicochemical properties, production technologies and applications[J]. Food Structure, 2017, 14: 17-29.
[3] 魏倩,张莹莹,阎欣,等. 豆类蛋白和谷物蛋白的功能性对比[J]. 粮食加工, 2018, 43(1): 36-41.
[4] 王盼盼. 食品中蛋白质的功能(三) 食品中蛋白质的功能特性[J]. 肉类研究, 2009(6): 71-77.
[5] BOWKER B C, GRANT A L, SWARTZ D R. Myosin heavy chain isoforms influence myofibrillar ATPase activity under simulated postmortem pH, calcium, and temperature conditions[J]. Meat Science, 2004, 67(1): 139-147.
[6] AROGUNDADE L A. Functional characterization of Tef (Eragostics tef) protein concentrate: Influence of altered chemical environment on its gelation, foaming, and water hydration properties[J]. Food Hydrocolloids, 2006, 20(6): 831-838.
[7] SUN Fengyuan, HUANG Qilin, HU Ting, et al. Effects and mechanism of modified starches on the gel properties of myofibrillar protein from grass carp [J]. International Journal of Biological Macromolecules, 2014, 64: 17-24.
[8] SUN Lijun, SUN Jiaojiao, PRIDHUVI THAVARAJ, et al. Effects of thinned young apple polyphenols on the quality of grass carp (Ctenopharyngodon idellus) surimi during cold storage[J]. Food Chemistry, 2017, 224: 372-381.
[9] RAWDKUENA S, BENJAKUL S. Whey protein concentrate: Autolysis inhibition and efiects on the gel properties of surimi prepared from tropical fish[J]. Food Chemistry, 2008, 106: 1 077-1 084.
[10] SUN L, SUN J, PRIDHUVI T, et al. Effects of thinned young apple polyphenols on the quality of grass carp (Ctenopharyngodon idellus) surimi during cold storage[J]. Food Chemistry, 2017, 224: 372-381.
[11] MARÍN D, ALEMÁN A, SÁNCHEZ-FAURE A, et al. Freeze-dried phosphatidylcholine liposomes encapsulating various antioxidant extracts from natural waste as functional ingredients in surimi gels[J]. Food Chemistry, 2018, 245: 525-535.
[12] BUAMARD N, BENJAKUL S. Improvement of gel properties of sardine (Sardinella albella) surimi using coconut husk extracts[J]. Food Hydrocolloids, 2015, 51: 146-155.
[13] VON STASZEWSKI M, JAGUS R J, PILOSOF A M R, et al. Pilosof.Influence of green tea polyphenols on the colloidal stability and gelation of WPC[J]. Food Hydrocolloids, 2011, 25(5): 1 077-1 084.
[14] PRIGENT S V E, VORAGEN A G J, VAN KONINGSVELD G A. Interactions between globular proteins and procyanidins of different degrees of polymerization[J]. Journal of Dairy Science, 2009, 92(12): 5 844-5 853.
[15] ALU'DATT M H, RABABAH T, ALLI I. Effect of phenolic compound removal on rheological, thermal and physico-chemical properties of soybean and flaxseed proteins[J]. Food Chemistry, 2014, 146: 608-613.
[16] BALANGE A K, BENJAKUL S. Cross-linking activity of oxidised tannic acid towards mackerel muscle proteins as affected by protein types and setting temperatures[J]. Food Chemistry, 2010, 120: 268-277.
[17] KROLL J, RAWEL H M. Reactions of plant phenols with myoglobin: Influence of chemical structure of the phenolic compounds[J]. Food Chemistry and Toxicology, 2001, 66 (1): 48-57.
[18] CAO Y, XIONG Y. Chlorogenic acid-mediated gel formation of oxidatively stressed myofibrillar protein[J]. Food Chemistry, 2015, 180: 235-243.
[19] KROLL J, RAWEL H M, ROHN S. Reactions of plant phenolics with food proteins and enzymes under special consider-ation of covalent bonds[J]. Food Sci. Technol. Res, 2003,9 (3): 205-218.
[20] OJHA H, MISHRA K, I. HASSAN M, et al. Spectroscopic and isothermal titration calorimetry studies of binding interaction of ferulic acid with bovine serum albumin[J]. Thermochimica Acta, 2012(548): 56-64.
[21] NAGY K, COMPONDU M C C, WILLIAMSON G. Non-covalent binding of proteins to polyphenols correlates with their amino acid sequence[J]. Food Chemistry, 2012, 132: 1 333-1 339.
[22] 徐洁琼,曾茂茂,秦昉,等. 热加工处理对β-乳球蛋白与茶多酚间相互作用的影响[J]. 食品与发酵工业, 2017, 43(8): 96-102.
[23] 王育信. 褐藻多酚的提取和鉴定及其对鱼糜凝胶物性的影响[D]. 上海:上海海洋大学,2018.
[24] CONNAN S, GOULARD F, STIGER V, et al. Interspecific and temporal variation in phlorotannin levels in an assemblage of brown algae[J]. Botanica Mar, 2004, 47: 410-416.
[25] RAWEL H M, KROLL J, HOHL U C. Model studies on reactions of plant phenols with whey proteins[J]. Molecular Nutrion, 2001, 45(2): 72-81.
[26] RAWEL H M, ROHN S, KRUSE H P, et al. Structural changes induced in bovine serum albumin by covalent attachment of chlorogenic acid[J]. Food Chemistry, 2002, 78: 443-455.
[27] CROWELL E A. Determination of alpha amino nitrogen in musts and wines by TNBS method [J]. Journal of Enology and Viticulture, 1985, 36(2): 175-177.
[28] BEVERIDGE T, TOMA S J, NAKAI S. Determination of SH- and SS-groups in some food proteins using Ellman’s reagent[J]. Journal of Food Science, 1974, 39: 49-51.
[29] GÓMEZ-GUILLÉN M C, BORDERÍAS A J, MONTERO P. Chemical interactions of nonmuscle proteins in the network of sardine(Sardine pilchardus) muscle gels[J]. Lebensmittel-wissenschaft Und-technologie-food Science and Technology, 1997, 30(6): 602-608.
[30] ALIZADEH-PASDAR N,LI-CHAN E C. Comparison of protein surface hydrophobicity measured at various pH values using three different fluorescent probes[J]. Journal of Agricultural and Food Chemistry, 2000, 48: 328-334.
[31] 曹艳芸. 乳清蛋白与多酚在中性pH条件下的相互作用对蛋白功能性质的影响研究[D]. 无锡:江南大学, 2017.
[32] TANAKA T, KOUNO I. Oxidation of tea catechins: Chemical structures and reaction mechanism[J]. Food Science and Technology Research, 2003,9(2): 128-133.
[33] KROLL J, RAWEL H M, ROHN S. Reactions of plant phenolics with food proteins and enzymes under special consider-ation of covalent bonds[J]. Food Science and Technology Research, 2003,9(3): 205-218.
[34] RAWEL H M, KROLL U J, HOHL C. Model studies on reactions of plant phenols with whey proteins[J]. Molecular Nutrion, 2001, 45(2): 72-81.
[35] RAWEL H M, KROLL J, ROHN S. Reactions of phenolic substances with lysozyme—physicochemical characterisation and proteolytic digestion of the derivatives[J]. Food Chemistry, 2001(72): 59-71.
[36] KANG J, LIU Y, XIE M. Interactions of human serum albumin with chlorogenic acid and ferulic acid[J]. Food Chemistry, 2017, 236: 114-118.
[37] CHU Q, BAO B, WU W. Mechanism of interaction between phenolic compounds and proteins based on non-covalent and covalent interactions[J]. Medicine Research, 2018, 2: 1-6.
[38] 赵新淮, 徐红华, 姜毓敏. 食品蛋白质-结构、性质与功能[M].北京: 科学出版社, 2009: 207.
[39] OJHA H, MISHRA K, HASSAN M I, et al. Spectroscopic and isother-mal titration calorimetry studies of binding interaction of ferulic acid with bovineserum albumin[J]. Thermochimica Acta, 2012,548: 56-64.
[40] 齐宝坤,赵城彬,江连洲,等. 大豆分离蛋白组成及二级结构对表面疏水性的影响[J]. 中国食品学报, 2018, 18(5): 288-293.
[41] RAWEL H M, KROLL J, RIESE B. Reactions of chlorogenic acid with lysozyme: physicochemical characterization and proteolytic digestion of the derivatives[J]. Sensory and Nutritive Qualities of Food, 2000, 65(6): 1 091-1 097.
[42] GÜLER ,VOROB’EV M M, VOGEL V, et al. Proteolytically-induced changes of secondary structural protein conformation of bovine serum albumin monitored by Fourier transform infrared (FT-IR) and UV-circular dichroism spectroscopy[J]. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2016, 161: 8-18.
[43] JIANG J, ZHANG Z, ZHAO J, et al. The effect of non-covalent interaction of chlorogenic acid with whey protein and casein on physicochemical and radical-scavenging activity of in vitro protein digests[J]. Food Chemistry, 2018, 268: 334-341.
[44] ALI M, HOMANN T, KHALIL M, et al. Milk whey protein modification by coffee-specific phenolics: Effect on structural and functional propertie[J]. Journal of Agricultural and Food Chemistry, 2013, 61(28): 6 911-6 920.
[45] KIM S J, USTUNOL Z. Thermal properties, heat sealability and seal attributes of whey protein isolate/lipid emulsion edible films[J]. Journal of Food Science, 2001, 66(7): 985-990.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司