构建基于姜黄素-Hg^(2+)复合物的荧光传感器检测脂肪酶活性  

A Fluorescence Sensor for Determination of Lipase Activity Based on Curcumin-Hg^(2+) Complex

作　　者：吴胜男 张浩 江凌[2] 霍峰蔚[3] 田丹碧 WU Sheng-Nan;ZHANG Hao;JIANG Ling;HUO Feng-Wei;TIAN Dan-Bi(College of Chemistry and Molecular Engineering,Nanjing Tech University,Nanjing 211816,China;College of Food Science and Light Industry,Nanjing Tech University,Nanjing 211816,China;Institute of Advanced Material,Nanjing Tech University,Nanjing 211816,China)

机构地区：[1]南京工业大学化学与分子工程学院,南京211816 [2]南京工业大学食品与轻工学院,南京211816 [3]南京工业大学先进材料研究院,南京211816

出　　处：《分析化学》2018年第10期1588-1594,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金联合基金项目(Nos.U1603112;21574065);杰出青年科学基金项目(No.21625401)资助~~

摘　　要：以吐温40-姜黄素复合物胶束为荧光信号探针,Hg^(2+)为猝灭剂,巯基乙酸甲酯为脂肪酶的底物,构建了检测脂肪酶活性的生物传感器。利用Hg^(2+)与姜黄素β-二酮结构结合,猝灭吐温40-姜黄素复合物胶束荧光;而脂肪酶的加入催化了巯基乙酸甲酯水解生成巯基乙酸,巯基乙酸夺取姜黄素-Hg^(2+)复合物上的Hg^(2+),吐温40-姜黄素复合物胶束的荧光恢复,荧光恢复强度与脂肪酶的活性相关。利用此荧光传感策略定量检测脂肪酶活性,在最佳实验条件下,荧光强度变化值与脂肪酶活性在2~80 U/mL范围内呈线性关系,检出限为0.01 U/mL。采用此方法对另外4种商业脂肪酶的活性进行了检测,实验结果与恒电位滴定法测定结果一致。本方法检测成本低,操作简单,灵敏度高,具有良好的实用性,可用于脂肪酶活性的高通量检测。A biosensor for detecting lipase activity was constructed by using the Tween 40-curcumin complex micelle as a fluorescence probe,Hg 2+as a quenching agent and methyl thioglycolate as the substrate of lipase.Hg^2+combined with theβ-diketone structure of curcumin,resulting in quenching of Tween 40-curcumin complex micelle fluorescence.However,upon the addition of lipase,lipase could catalyze the hydrolysis of methyl thioglycolate to produce mercaptoacetic acid which could capture Hg^2+released from the curcumin-Hg^2+complex,resulting in the recovery of fluorescence of the Tween 40-curcumin complex micelle.The intensity of fluorescence recovery was related to lipase activity.The fluorescence sensing strategy was applied to quantitatively detect lipase activity.Under optimal experimental conditions,the change of fluorescence intensity was linear with the concentration of lipase in the range of 2-80 U/mL,and the detection limit was 0.01 U/mL.This method was employed to detect the activity of four other kinds of commercial lipases.The results were in agreement with those determined by potentiometric titration.The results showed that the method had the advantages of low cost,simple operation,high sensitivity,good practicability,and could be used for high throughput detection of lipase activity.

关 键 词：脂肪酶活性 吐温40-姜黄素复合物 汞离子 巯基乙酸甲酯 荧光传感 

分 类 号：O657.3[理学—分析化学]