大豆11S球蛋白糖基化有害交联产物的形成及抑制机理  

Formation and Inhibition Mechanism of Harmful Cross-Linked Products Generated from Glycosylation of 11S Globulin

作　　者：章鼎敏 刘贵梅[1] 卢永翎[1] 郑铁松[1] 吕丽爽[1] ZHANG Dingmin;LIU Guimei;LU Yongling;ZHENG Tiesong;LüLishuang(Ginling College,Nanjing Normal University,Nanjing 210097,China)

机构地区：[1]南京师范大学金陵女子学院,江苏南京210097

出　　处：《食品科学》2018年第7期7-13,共7页Food Science

基　　金：国家自然科学基金面上项目(31571783);江苏省研究生科研与实践创新计划项目(KYCX17-1089)

摘　　要：目的:针对食品工业大豆蛋白糖基化改性工艺,建立大豆11S球蛋白-糖模拟体系,探究该体系中晚期糖基化终产物(advanced glycation end products,AGEs)形成的影响因素,探究黄酮类化合物抑制AGEs形成的机理,以便进行有效调控。方法:荧光光谱法(λ_(ex)/λ_(em)=340 nm/465 nm)测定糖的种类和浓度、温度、pH值以及黄酮抑制剂等对AGEs形成的影响,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表征糖基化过程及黄酮对AGEs的抑制效果,通过液相色谱-串联质谱(liquid chromatograph-tandem mass spectrometry,LC-MS/MS)测定反应过程中槲皮素及其产物的变化。结果:糖种类为果糖;11S球蛋白与果糖质量浓度分别为2、8 mg/m L,pH值为9.2,温度为121℃条件下产生的荧光性AGEs量最多;槲皮素、木犀草素、染料木素、芦丁4种黄酮对荧光性AGEs的形成均有一定抑制效果;LC-MS/MS结果表明槲皮素通过捕获糖基化中间产物甲基乙二醛(methylglyoxal,MGO),抑制了大分子交联性的荧光性AGEs形成;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳验证了1 mmol/L槲皮素有效降低了大分子交联的产物,但不影响糖的修饰改性,乳化性能得到较好的改善。结论:采用分子质量较大的糖,降低糖质量浓度、pH值和反应温度,添加黄酮类化合物可有效抑制大豆蛋白糖基化过程中有害产物AGEs的形成。Purpose:The objective of this research is to identify the factors affecting the formation of advanced glycation end products(AGEs)in 11S globulin-sugar model system and to shed light on the inhibitory mechanism of flavonoids on AGEs formation.Methods:Fluorescence spectrometry(λex/λem=340 nm/465 nm)was used to detect the effects of sugar type and concentration,temperature,pH and flavonoids on the formation of AGEs,and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)was employed to characterize the glycation process and investigate the inhibitory effect of flavonoids.Changes in the concentrations of quercetin and products during the reaction process were determined by liquid chromatograph-tandem mass spectrometry(LC-MS/MS).Results:The reaction between 11S globulin at 2 mg/mL and fructose at 8 mg/mL at pH 9.2,121℃formed the greatest amount of fluorescent AGEs.The formation of AGEs was inhibited by all four flavonoids(quercetin,luteolin,genistein,or rutin).LC-MS/MS analysis showed that quercetin inhibited the generation of fluorescent cross-linked AGEs by trapping methylglyoxal as an intermediate of glycolysis.SDS-PAGE analysis confirmed that quercetin at 1 mmol/L significantly reduced the generation of cross-linked macromolecules but did not influence sugar modification and resulted in significant improvement of emulsifying properties.Conclusion:Application of high-molecular weight sugars,reduced sugar concentration,pH and reaction temperature,and addition of flavonoids can effectively inhibit the formation of harmful products AGEs during the glycosylation process of soybean protein.

关 键 词：大豆11S球蛋白 晚期糖基化终产物 黄酮 蛋白糖基化改性 

分 类 号：TS201.2[轻工技术与工程—食品科学]