大豆蛋白金纳米簇“关—开”型荧光探针检测炭疽杆菌生物标志物DPA  

Soy Protein Gold Nanocluster as an “Off-On” Fluorescent Probe for the Detection of Bacillus Anthracis Biomarkers DPA

作　　者：钱多 苏文恩 刘致远 高小钰 易雨欣 胡聪聪 刘斌 杨胜园[1] QIAN Duo;SU Wen-en;LIU Zhi-yuan;GAO Xiao-yu;YI Yu-xin;HU Cong-cong;LIU Bin;YANG Sheng-yuan(School of Public Health,University of South China,Hengyang 421001,China)

机构地区：[1]南华大学公共卫生学院,湖南衡阳421001

出　　处：《光谱学与光谱分析》2023年第6期1815-1820,共6页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(82073604);湖南省研究生科研创新项目(CX20210962);国家级大学生创新创业训练计划项目(202110555008)资助。

摘　　要：炭疽杆菌是高致病性病原微生物,引起的炭疽病在我国属于乙类传染病,因此建立操作简便、灵敏准确的炭疽杆菌检测方法对预防和控制炭疽传播,维护公共卫生安全至关重要。该研究创新性地提出以绿色材料大豆蛋白为保护剂和还原剂,采用微波加热法合成了一种具有强烈红色荧光发射的大豆蛋白金纳米簇(SPI-AuNCs)。采用TEM、 UV-Vis、 FL、 XPS、 FTIR等方法表征了SPI-AuNCs的成功合成和部分特殊性能。结果表明SPI-AuNCs呈球形,粒径大小在1.8~3.2 nm范围内,平均粒径为2.65 nm,在500~550 nm范围内未出现表面等离子体共振峰;SPI-AuNCs的最大激发波长为370 nm,最大发射波长为680 nm。SPI-AuNCs的表面官能团主要有—NH、—COOH、—OH、—SH等官能团,元素的组成主要包括C、 N、 O、 S、 Au元素。根据Cu^(2+)与SPI-AuNCs表面基团通过配位作用形成非荧光复合物,使荧光猝灭;而2,6-吡啶二羧酸(DPA)与Cu^(2+)具有更高的亲和力,可将Cu^(2+)从SPI-AuNCs表面竞争下来,使荧光恢复,据此建立了一种荧光“关-开”策略检测DPA的新方法。在最佳实验条件下,荧光恢复率(ΔF/F_(1))与DPA的浓度在1.15~70.0μmol·L^(-1)范围内呈良好的线性关系,线性方程为ΔF/F_(1)=0.011c+0.131,相关系数r=0.991,方法检出限为0.34μmol·L^(-1)。同时,通过加标回收实验研究了湖水和牛奶样品中DPA,得到加标回收率在97.3%~103.6%,表明了该方法在环境和食品样本DPA的检测中具有很大的应用潜力,可以为环境监测和食品安全提供方法学支持。Bacillus anthracis is a highly pathogenic microorganism.Anthrax is an infectious disease caused by Bacillus anthracis,classified as Class B of our country’s statutory reporting infectious diseases.Therefore,the establishment of simple operation,rapid and sensitive detection methods for Bacillus anthracis is vital for preventing and contralling the spread of anthrax and maintaining public health security.This study innovatively proposed to synthesize soy protein gold nanoclusters(SPI-AuNCs)with strong red fluorescence emission by microwave heating using the green material soy protein as a protective agent and reducing agent.TEM,XPS,FTIR,FL,and UV-Vis characterizations were used to verify successful synthesis of SPI-AuNCs.SPI-AuNCs are spherical,which the particle size is in the range of 1.8~3.2 nm,an average diameter of 2.65 nm,and no surface plasma resonance absorption from 500 to 550 nm.The maximum excitation wavelength of SPI-AuNCs is 370 nm,and the maximum emission wavelength appears at 680 nm.The surface functional groups of SPI-AuNCs mainly include—NH,—COOH,—OH,—SH,and the element composition includes C,N,O,S,Au elements.The fluorescence of SPI-AuNCs could be quenched by the coordination between Cu^(2+)and surface groups of SPI-AuNCs,while DPA has a stronger chelation effect with Cu^(2+),which could compete for Cu^(2+)from the surface of SPI-AuNCs and restore the fluorescence of SPI-AuNCs.Accordingly,a new method for DPA detection based on the fluorescent“off-on”strategy was established.Under the optimal experimental conditions,the fluorescence recovery efficiency(ΔF/F 1)performs a good linear relationship with DPA concentration in the range of 1.15~70.0μmol·L^(-1).The linear regression equation isΔF/F 1=0.011 c+0.131 with high correlation coefficient(r=0.991),and the detection limit is 0.34μmol·L^(-1).In addition,the spiking experiments of the DPA in lake water and milk samples were performed.The spiked recoveries were 97.3%~103.6%,indicating that this method has great application potent

关 键 词：大豆蛋白金纳米簇 2 6-吡啶二羧酸 生物标志物 炭疽杆菌 荧光探针 

分 类 号：O657.3[理学—分析化学]