超声协同糖基化对蛋清粉致敏性及结构的影响

以市售蛋清粉为研究对象,通过电泳、紫外光谱、荧光光谱和酶联免疫吸附等方法研究超声波协同糖基化修饰对其蛋白结构和致敏性的影响。结果表明,超声波协同糖基化修饰能显著降低蛋清粉的致敏性,致敏性随超声强度的增加呈先增加后降低的趋势,且在15 min、600 W的条件下有最大程度降低。这和其结构变化密切相关,经复合处理后,蛋清蛋白分子量显著增加,同时三级结构发生显著变化,使表面疏水性增强、内源荧光和自由氨基含量降低,导致其致敏性降低。超声波协同糖基化修饰是一种良好的降低蛋清粉致敏性的方法。     

美拉德反应对产品风味品质的影响及其衍生危害物研究进展

在食品加工过程中,美拉德反应虽然对提升食品的风味和品质具有重要作用,但也会衍生一些化学危害物。本文综述了美拉德反应对食品风味和色泽等品质的影响,以及美拉德反应产物的抗氧化和抗突变等特性,同时也探讨了美拉德反应产生的丙烯酰胺(acrylamide,AA)、杂环胺(heterocyclic amines,HAs)和晚期糖基化末端终产物(advanced glycation end products,AGEs)等化学危害物的生成机制、分析和抑制方法。由于3类危害物在食品中的含量较低(ng/g~μg/g)且基质非常复杂,因此其检测均需要采用固相萃取等方法进行分离富集,随后可采用超高效液相色谱串联质谱(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)等方法进行定性定量分析。危害物的抑制主要通过采用更加温和的加工方法、调整工艺参数和添加香辛料及酚类等天然抗氧化物质来实现。     

Strategies for proteomic analysis of non-enzymatically glycated proteins

Among post-translational modifications of proteins, non-enzymatic glycation is one of the less frequently studied by experts in proteomics. However, the relevance of protein glycation has been widely shown up in several pathological conditions. In fact, non-enzymatic glycation has been strongly related to hyperglycemic conditions and, thus, to chronic complications associated to diabetes mellitus and renal failure as well as degenerative changes occurring in the course of aging. Two different glycation levels are distinguished whether the structure of the protein is seriously damaged or not. The biochemical and clinical significance of both glycations have been already described. Several reasons have contributed to the lack of highly sensitive and selective methods for identification and quantitation of glycated proteins. These are mainly (1) the low concentration of glycated proteins in humans due to the low efficiency of the glycation process, (2) the modification of enzymatic digestion patterns, (3) the low ionization efficiency of glycated peptides, and (4) the lack of software including tools to identify this post-translational modification. The aim of this review is to provide the analytical guidelines required to succeed in the analysis of glycated proteins. For this purpose, different analytical approaches are considered to solve the main drawbacks derived from this gap in the proteomics field. Some challenges are finally proposed to be taken into account in future research.     

糖基化反应改善鱼肉肌原纤维蛋白功能特性及其机制研究进展

糖基化反应是一种有效改善蛋白质功能特性的方法。文中结合国内外有关鱼肉肌原纤维蛋白糖基化改性的最新研究进展,综述了糖基化改性对于鱼肉肌原纤维蛋白功能特性的影响,探讨了鱼肉肌原纤维蛋白—多糖复合物的形成及其功能特性变化的机理。     

桑椹花色苷对晚期糖基化末端产物抑制作用及其机制分析

晚期糖基化末端产物(Advanced Glycation End Products,AGEs)是蛋白质游离氨基与还原糖羰基反应形成的一类复杂物质的总称,是诱发糖尿病等多种慢性病的重要因素之一,而多酚类化合物能够抑制AGEs的形成。为了探讨桑椹花色苷对AGEs的抑制机制,建立牛血清蛋白(BSA)/葡萄糖(Glucose)生成AGEs模拟反应体系,以0.5%酸性乙醇提取、AB-8型大孔树脂纯化的桑椹花色苷为抑制剂,测定反应体系中蛋白质羰基、巯基含量变化、羟基自由基(·OH)对AGEs形成的影响,利用SDS-PAGE凝胶电泳技术分析BSA的分子量变化。研究结果表明,随着桑椹花色苷浓度的增加,反应体系中的AGEs含量显著降低(P0.05)。桑椹花色苷可显著抑制·OH诱导的AGEs的形成(P0.05)。桑椹花色苷通过保护巯基和抑制蛋白质形成羰基而抑制AGEs生成;SDS-PAGE图谱显示,桑椹花色苷可阻断糖基化反应导致的蛋白质交联物的形成。     

油炸方便面贮藏过程中糖基化产物的变化规律

油炸食品贮藏过程中易形成乙二醛(glyoxal,GO)、丙酮醛(methylglyoxal,MGO)等糖基化中间产物,进一步反应生成晚期糖基化终产物(advanced glycation end prodcts,AGEs),包括荧光性AGEs和非荧光性AGEs,而AGEs与人类诸多疾病的发生密切相关。本试验研究了油炸方便面在贮藏过程中,温度、氧气、光照对GO、MGO、荧光性AGEs变化的影响。结果表明,温度可促进GO、MGO的变化,随时间延长呈现先上升后降低的趋势。氧气能促进GO含量的降低,对MGO变化没有显著性影响。光照对GO、MGO的变化无显著性影响;随着贮藏时间延长,荧光性AGEs含量在不同贮藏条件下都有所增加,其中温度和氧气的影响较大,37℃和室温有氧条件分别达到342.17、340.15 AU/g。因此,油炸方便面贮藏时应低温、密封、避光,以保证其品质,减少糖基化产物的形成。     

Potential DNA–protein cross-link products formed by sugar degradation products: identification of N 6-[2-(N 2-2′-deoxyguanosyl)propionyl]lysine

Sugar degradation products are formed during heat treatment of food as well as endogenously in vivo. As reactive carbonyl compounds, they react readily with proteins or DNA to form protein- or DNA-bound advanced glycation end products (glycation reaction or Maillard reaction). In this study, we investigated the formation of potential DNA–protein cross-link products from sugar degradation products. 2-Deoxyguanosine, l-lysine and different carbohydrates were incubated at 37 °C. The sugar degradation products dihydroxyacetone and d,l-glyceraldehyde lead to the formation of two new cross-link products. The new compounds were isolated by preparative high-performance liquid chromatography and identified by spectral data as the two diastereomers of N⁶-[2-(N²-2-deoxyguanosyl)propionyl]lysine. In this structure, the -amino group of lysine and the exocyclic amine group of 2-deoxyguanosine are linked via a carboxyethyl group, derived from the carbohydrate component. The binding sites and the binding types were confirmed by synthesis of the analogous products from N²-(1-carboxyethyl)guanosine and N-acetyllysine methyl ester.Dedicated to Professor Th. Severin on the occasion of his 75^th birthday     

超声波辅助糖基化对β-乳球蛋白消化过程中致敏性的影响

采用间接竞争ELISA、扫描电镜和动态光散射等技术,研究超声波结合乳糖糖基化修饰对β-乳球蛋白(β-Lg)在体外模拟消化过程中致敏性和结构变化的影响。结果表明:被修饰的β-Lg在消化过程中致敏性显著降低,胃肠消化的水解度也降低;被修饰的的β-Lg经胃消化形成更大的颗粒结构,在肠消化中发生聚集;消化后其荧光强度先升高后降低,最大发射波长出现红移。综上,超声波结合糖基化改性可有效降低β-Lg在消化过程中的致敏性,其致敏性变化与糖基化修饰对其结构的改变有关。