超声波处理对绿原酸与牛血清白蛋白结合作用的影响  

Effect of Ultrasonic Treatment on the Binding of Chlorogenic Acid to Bovine Serum Albumine

作　　者：范金波 麻奥 闵爽 邢莉 吕长鑫 励建荣 Fan Jinbo;Ma Ao;Min Shuang;Xing Li;LüChangxin;Li Jianrong(College of Food Science and Technology,Bohai University,Food Safety Key Lab of Liaoning Province,National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,Jinzhou 121013,Liaoning)

机构地区：[1]渤海大学食品科学与工程学院,辽宁省食品安全重点实验室,生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心,辽宁锦州121013

出　　处：《中国食品学报》2022年第3期129-138,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金面上项目(31972107)。

摘　　要：采用荧光光谱法和紫外-可见吸收光谱法研究超声处理(40 kHz/30 min和50 kHz/30 min)对绿原酸(CGA)与牛血清白蛋白(BSA)作用的影响。通过荧光发射光谱来分析CGA对BSA的荧光猝灭作用;通过同步荧光光谱分析结合CGA后BSA酪氨酸(Tyr)残基与色氨酸(Trp)残基微环境的变化来确定CGA对BSA构象的影响;运用Stern-Volmer方程、Acharya方程等数学方程计算其猝灭常数、结合常数,以及结合位点数;通过焓变与熵变分析分子间作用力。利用华法林(WARF)、布洛芬(IBU)两种蛋白标记物来判断结合位点;利用紫外光谱测定BSA构象变化对比不同超声频率的影响。结果表明:在超声频率53 kHz时,CGA对BSA的荧光猝灭效果最为显著,超声处理使CGA-BSA复合物的构象发生一定改变,并对CGA与BSA的结合有增强作用,而不会改变反应的作用力类型。研究结果为超声处理对蛋白质与配体结合的影响研究提供参考。The paper uses fluorescence spectroscopy and ultraviolet-visible absorption spectroscopy to study the effect of ultrasonic treatment(40 kHz/30 min,50 kHz/30 min)on the binding of chlorogenic acid(CGA)and bovine serum albumin(BSA).Analyze the fluorescence quenching effect of CGA on BSA by fluorescence emission spectroscopy;analyze the microenvironmental changes of BSA tyrosine(Tyr)and tryptophan(Trp)residues after CGA binding by synchronous fluorescence spectroscopy to determine the effect of CGA on the conformation of BSA;use mathematical equations such as Stern-Volmer equation and Acharya equation to calculate its quenching constant,binding constant,and number of binding sites;then use warfarin(WARF)and ibuprofen(ibuprofen,IBU)two protein markers to determine the binding site;and use ultraviolet spectroscopy to measure the changes in BSA conformation to compare the effects of different ultrasonic frequencies.The results showed that CGA had the most significant fluorescence quenching effect on BSA at the ultrasonic frequency of 53 kHz,and the conformation of CGA-BSA complex could be changed by ultrasonic treatment to some extent,and the combination of CGA and BSA could be enhanced to some extent,but the reaction force type would not be changed.The results of this study will provide a useful reference for the effect of sonication on the binding of protein and ligand.

关 键 词：牛血清白蛋白 绿原酸 超声波处理 荧光猝灭 紫外-可见吸收光谱法 

分 类 号：TB559[理学—物理] TS201[理学—声学] O657.3[轻工技术与工程—食品科学]