食品体系中糖基化蛋白质结构的表征和检测方法综述

doi:10.3964/j.issn.1000-0593(2023)09-2667-07

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摘要：蛋白质和糖是食品的重要组分。在食品加工、贮运过程中，蛋白质分子中的氨基与还原糖的羰基很容易以共价键的形式相结合发生糖基化反应。美拉德式糖基化反应包括早期、中期和末期三个阶段。糖基化反应使得蛋白质分子主链或侧链发生修饰，改变蛋白质结构、静电荷和疏水性等，影响蛋白质主链和侧链基团的化学活性，进而改变蛋白质的理化功能性质，如改善乳化性、起泡性、凝胶性等功能性质，提高抗氧化性等营养特性，降低致敏性，增强抑菌性和贮藏特性等，在食品加工、贮运过程中非常普遍且有着重要意义。糖基化反应过程中引起的蛋白质结构变化是其营养、功能特性改变的重要原因。近年来，化学测定（自由氨基含量、表面疏水性测定等）、光谱（紫外、荧光、傅里叶变换红外光谱等）、色谱（圆二、分子排阻色谱等）、质谱[基质辅助激光解吸附离子化-飞行时间质谱（MALDI-TOF-MS）、液相色谱-电喷雾串联质谱（LC-MS²）等]等多种技术手段被探索用于分析蛋白质糖基化反应过程中的结构变化规律，在糖基化反应机制研究及糖基化反应程度、产物营养功能性质调控等方面起了重要作用。尤其是轨道离子阱质谱（LC-Orbitrap-MS²）、傅里叶变换离子回旋共振质谱（LC-FTICR-MS²）、氢氘交换-傅里叶变换离子回旋共振质谱（HDX-FTICR-MS²）等技术的出现和成熟应用让糖基化位点研究更加深入，可以对各位点的糖分子连接数和糖基化取代程度进行定量分析，使得揭示蛋白质糖基化反应机制变为可能。对基于蛋白质糖基化反应程度、一级、二级、三级结构和官能团结构的表征和检测方法进行综述，旨在为蛋白质糖基化反应的深入研究和在食品加工中的应用提供参考。

关键词：食品；蛋白质；糖基化；结构；表征；检测方法

Abstract：Proteins and sugars are important components of food. During food processing, storage and transportation, the amino group of protein molecules is easily combined with the carbonyl group of reducing sugar through covalent bonds, and the glycation reaction occurs. Maillard reaction-based glycation includes three stages: the early stage, the middle stage and the advanced stage. The glycation reaction modifies the main or side chains of proteins, changes the structure, static charge and hydrophobicity of proteins, and affects the chemical activities of the main and side chain groups of proteins, thus changing the physicochemical properties of proteins, including improving the emulsifying, foaming, gel and other functional properties, enhancing the antioxidant and other nutritional properties, while reducing the sensitization and enhancing the antibacterial and storage properties. It is very common and important in food processing, storage and transportation. The changes in protein structure during the reaction are the important causes of the alteration of nutritional and functional properties. In recent years, chemical analysis (free amino content and surface hydrophobicity determination, etc.), spectrometry (ultraviolet spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, etc.), chromatography (circular dichroism, size-exclusion high-performance liquid chromatography, etc.), mass spectrometry (matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), liquid chromatography-electrospray tandem mass spectrometry (LC-MS²), etc.) and other techniques have been explored to analyze the structural changing rules of protein during glycation reaction, playing the important roles in the mechanism study of glycation reaction as well as regulation of glycation degree and nutritional and functional properties of glycation products. In particular, the emergence and mature application of orbital ion trap mass spectrometry (LC-Orbitrap-MS²), Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS²), hydrogen-deuterium exchange Fourier transform ion cyclotron resonance mass spectrometry (HDX-FTICR-MS²), and other technologies have made the research on glycation sites more in-depth. These technologies make it possible to reveal the mechanism of protein glycation reaction by quantifying the number of bonded sugar molecules and the degree of glycation substitution at each reaction site. This study reviewed characterization and detection methods for the protein glycation degree, primary, secondary and tertiary protein structures and functional group structure. This study aims to provide a reference for the deep research of protein glycation reaction and its application in food processing.

Key words：Food; Protein; Glycation; Structure; Characterization; Detection methods

收稿日期: 2022-05-07 修订日期: 2022-07-26

中图分类号:

TS201.21

基金资助: 国家自然科学基金项目（32101946），江西省自然科学基金项目（20212BAB215037）资助

通讯作者: 胡月明 E-mail: huyueming@ncu.edu.cn

作者简介: 张鹏，1977年生，江西福美泰生物技术有限公司硕士研究生 e-mail: lemon@foodchem.cn

引用本文:

张鹏，杨一帆，王辉，涂宗财，沙小梅，胡月明. 食品体系中糖基化蛋白质结构的表征和检测方法综述[J]. 光谱学与光谱分析, 2023, 43(09): 2667-2673.
ZHANG Peng, YANG Yi-fan, WANG Hui, TU Zong-cai, SHA Xiao-mei, HU Yue-ming. A Review of Structural Characterization and Detection Methods of Glycated Proteins in Food Systems. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(09): 2667-2673.

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