食品科学 ›› 2018, Vol. 39 ›› Issue (7): 7-13.doi: 10.7506/spkx1002-6630-201807002

• 基础研究 • 上一篇    下一篇

大豆11S球蛋白糖基化有害交联产物的形成及抑制机理

章鼎敏,刘贵梅,卢永翎,郑铁松,吕丽爽*   

  1. 南京师范大学金陵女子学院,江苏 南京 210097
  • 出版日期:2018-04-15 发布日期:2018-04-17
  • 基金资助:
    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)

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

ZHANG Dingmin, LIU Guimei, LU Yongling, ZHENG Tiesong, Lü Lishuang*   

  1. Ginling College, Nanjing Normal University, Nanjing 210097, China
  • Online:2018-04-15 Published:2018-04-17

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(λex/λem=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/mL,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。

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

Abstract: 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.

Key words: 11S globulin, advanced glycation end products, flavonoids, protein glycosylation modification

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