基于植物酯酶-Cu_(3)(PO_(4))_(2)杂化纳米花的酶抑制型方法检测敌敌畏残留  

Detection of Dichlorvos Residue by Enzyme Inhibition Method Based on Plant Esterase-Cu_(3)(PO_(4))_(2) Hybrid Nanoflowers

作　　者：曲林姣 王金鑫 姜磊[1] 马洪超[1] 杨丽敏[1] QU Lin-Jiao;WANG Jin-Xin;JIANG Lei;MA Hong-Chao;YANG Li-Min(Center for Bioengineering and Biotechnology,China University of Petroleum(East China),Qingdao 266580,China)

机构地区：[1]中国石油大学(华东)生物工程与技术中心,青岛266580

出　　处：《分析化学》2021年第9期1506-1514,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.31871878);中国石油大学(华东)大学生创新创业训练计划项目(No.202003022)资助。

摘　　要：制备了植物酯酶-Cu_(3)(PO_(4))_(2)杂化纳米花,实现了植物酯酶的固定,保持了其水解酶活性,而且引入的磷酸铜骨架不仅具有支撑作用,还具有过氧化物酶模拟酶活性。与游离酶不同,杂化纳米花具有较高的稳定性,在储存30天后,其水解酶活性和过氧化物酶模拟酶活性都维持在80%以上;同时,分层纳米结构具有较大的比表面积,有利于酶与底物之间的反应。基于此杂化纳米花的双酶活性,将其运用于1-萘酚偶联双酶反应的酶抑制型检测方法,实现了有机磷农药敌敌畏的灵敏检测。本方法对敌敌畏的检出限(LOD)为0.26 pmol/L,实际蔬菜水果样品中敌敌畏的回收率为96.8%~107.4%。Plant esterase-Cu_(3)(PO_(4))_(2)hybrid nanoflowers were prepared,which not only achieved the immobilization of plant esterase and maintained its hydrolase activity,but also skillfully used copper phosphate skeleton.The copper phosphate skeleton not only had the supporting role,but also had the peroxidase-like activity.Different from free enzymes,hybrid nanoflowers showed higher stability,and their hydrolase activity and peroxidase-like activity maintained above 80%after storage for 30 days.Meanwhile,the layered nanostructures had larger specific surface area,which facilitated the reaction between enzyme and substrate.Due to the dual enzyme activity,the hybrid nanoflowers were applied to the enzyme inhibition-based colorimetric assay of 1-naphthol-linked bienzymatic reaction for the sensitive detection of organophosphorus pesticides.The limit of detection(LOD)for dichlorvos was as low as 0.26 pmol/L.The recoveries of dichlorvos in vegetables and fruits were 96.8%-107.4%.

关 键 词：植物酯酶-Cu_(3)(PO_(4))_(2)杂化纳米花 水解酶活性 过氧化物酶模拟酶活性 酶抑制型检测 有机磷农药 敌敌畏 

分 类 号：TS255.7[轻工技术与工程—农产品加工及贮藏工程]