紫外和荧光光谱法研究油菜蜂花粉多酚对胰脂肪酶的作用过程  

Study on the Interaction Process between Rape Bee Pollen Polyphenols and Pancreatic Lipase by Ultraviolet and Fluorescence Spectroscopy

作　　者：张楠 胡童霞 朱鑫麗 杨雪梅 王磊 吴帆 李红亮 ZHANG Nan;HU Tongxia;ZHU Xinli;YANG Xuemei;WANG Lei;WU Fan;LI Hongliang(College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Zhejiang Sunshine Analysis and Testing Co.,Ltd.,Hangzhou 311519,China)

机构地区：[1]中国计量大学生命科学学院,浙江杭州310018 [2]浙江分阳检测有限公司,浙江杭州311519

出　　处：《食品工业科技》2023年第22期36-42,共7页Science and Technology of Food Industry

基　　金：浙江省基础公益研究计划项目(LGN22C170001);国家自然科学基金(32170531);浙江省大学生科技创新活动计划暨新苗人才计划项目(2021R409049)。

摘　　要：本文利用酶动力学方法测定了油菜蜂花粉多酚对胰脂肪酶的抑制作用,并通过紫外光谱、荧光光谱和同步荧光光谱分析了油菜蜂花粉多酚与胰脂肪酶的相互作用过程。结果表明,油菜蜂花粉多酚对胰脂肪酶的活性具有一定的抑制作用,其半抑制浓度(IC_(50))为1.670±0.045 mg/mL,抑制类型为可逆性抑制中的混合型抑制。荧光光谱分析显示,随着油菜蜂花粉多酚浓度增加,胰脂肪酶的最大荧光发射强度逐渐降低,同时荧光最大发射波长从341 nm红移到349 nm。同步荧光光谱表明,多酚与胰脂肪酶结合时酶内部的疏水性降低,肽链的伸展程度增加;荧光猝灭机理结果显示,猝灭常数随温度升高从0.1000减小到0.0743,其结合过程为静态猝灭,且结合位点约为1个。综上,油菜蜂花粉多酚通过改变胰脂肪酶构象来抑制胰脂肪酶的活性,这为深入研究蜂花粉减脂机理提供了一定理论依据。In this study,the inhibitory effect of rape bee pollen polyphenols on pancreatic lipase was measured by enzyme kinetics method,and the interaction between pancreatic lipase and rape bee pollen polyphenol extracts was analyzed by the ultravilolet,fluorescence spectra and synchronous fluorescence spectra.The results showed that rape bee pollen polyphenols had a certain inhibitory effect on the activity of pancreatic lipase,and the half maximal inhibitory concentration(IC_(50))was 1.670±0.045 mg/mL.The inhibitory type was a mixed inhibition of reversible inhibition.Fluorescence spectra showed that the maximal fluorescence intensity of pancreatic lipase gradually decreased with the increasing of rape bee pollen polyphenols concentration,and the maximal fluorescence peak had a redshift from 341 nm to 349 nm.Synchronous fluorescence spectra showed that the hydrophobicity inside the enzyme decreased and the stretch of the peptide chain increased when polyphenols bind to pancreatic lipase.The fluorescence quenching mechanism showed that the quenching constant decreased from 0.1000 to 0.0743 with increasing temperature,and the binding process was static quenching with about one binding site.In conclusion,the rape bee pollen polyphenols could inhibit the pancreatic lipase activity by changing the enzyme conformation,which provided a certain theoretical basis for the mechanism of bee pollen lipid reduction.

关 键 词：油菜蜂花粉 多酚 胰脂肪酶 荧光光谱 抑制作用 

分 类 号：TS201.2[轻工技术与工程—食品科学]