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
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  • 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

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
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  • 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

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。

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.

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