摘要
以牛α-乳白蛋白(bovineα-lactalbumin,α-La)和果糖为非酶糖基化模型,分析槲皮素-3-O-葡萄糖苷(quercetin-3-O-β-D-glucuronide,Q3G)对α-La非酶糖基化过程中果糖胺和5-羟甲基糠醛(5-hydroxymethylfurfural,5-HMF)形成及游离氨基含量和褐变度的影响,然后结合分子对接和质谱鉴定技术进一步揭示其对非酶糖基化的抑制机理。结果表明:Q3G能显著减轻糖基化过程中的褐变程度,增加游离氨基含量,抑制果糖胺和5-HMF的形成,延缓糖基化反应的进程,且Q3G添加量为2 mmol/L时效果最好。分子对接结果表明Q3G可通过氢键、范德华力和疏水相互作用与α-La的活性氨基酸残基进行结合,影响α-La和果糖的糖基化反应,缓解糖基化反应的进程。同时,超高效液相色谱-四极杆-静电场轨道离子阱串联质谱分析也发现Q3G屏蔽了糖基化的潜在活性位点Lys93和Lys108,且新增糖基化位点Lys98的糖基化活性较弱,可达到抑制非酶糖基化的作用。本研究可为Q3G作为潜在抗糖基化剂的开发提供理论基础。
In this study,the effect of quercetin-3-O-β-D-glucuronide(Q3G)on the formation of fructosamine and 5-hydroxymethylfurfural(5-HMF),the amount of free amino groups and browning degree in a non-enzymatic glycosylation model consisting of bovineα-lactalbumin(α-La)and fructose were evaluated.In addition,the inhibitory mechanism of Q3G against non-enzymatic glycosylation was explored by molecular docking and mass spectrometry(MS).The results indicated that Q3G could significantly reduce the browning degree,increase the free amino groups,inhibit the formation of fructosamine and 5-HMF,and delay the process of glycosylation reaction.The effect was most pronounced at 2 mmol/L Q3G.Molecular docking analysis indicated that Q3G could bind with the active amino acid residues ofα-La through hydrogen bonds,van der Waals force and hydrophobic interactions,delaying the process of glycosylation reaction.Meanwhile,by ultra-high performance liquid chromatography-quadrupole-orbitrap tandem mass spectrometry(UPLC-Q-Orbitrap MS/MS)analysis,it was found that Q3G shielded the potential glycosylation sites Lys93 and Lys108,and the activity of the glycosylation site Lys98 was weak,indicating inhibited non-enzymatic glycosylation reaction.This study can provide a theoretical basis for the development of Q3G as a potential anti-glycosylation agent.
作者
张露
徐林菊
彭春彦
王佩欣
谢作桦
谢星
贾晓燕
涂宗财
ZHANG Lu;XU Linju;PENG Chunyan;WANG Peixin;XIE Zuohua;XIE Xing;JIA Xiaoyan;TU Zongcai(National R&D Center for Freshwater Fish Processing,College of Life Sciences,Jiangxi Normal University,Nanchang 330022,China;Jiangxi Deshang Pharmaceutical Co.Ltd.,Zhangshu 331200,China;State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2023年第20期28-34,共7页
Food Science
基金
国家自然科学基金地区科学基金项目(31860475)
江西省自然科学基金面上项目(20212BAB205017)。
关键词
槲皮素-3-O-葡萄糖苷
Α-乳白蛋白
分子对接
糖化位点鉴定
非酶糖基化
quercetin-3-O-β-D-glucuronide
bovineα-lactalbumin
molecular docking
identification of glycosylation sites
nonenzymatic glycosylation
