title: Fluorescence biosensor for inorganic pyrophosphatase activity.
abstract: A highly sensitive and selective fluorescence biosensor for inorganic pyrophosphatase (PPase) activity has been developed based on special click ligation trigger hyperbranched rolling circle amplification (CLT-HRCA). Pyrophosphate ion (PPi) can coordinate with Cu2+to form stable PPi/Cu2+complex and Cu2+in the complex cannot be reduced to Cu+. The addition of PPase causes the hydrolysis of PPi into orthophosphate (Pi) and therefore induces the releasing of Cu2+from the stable PPi/Cu2+complex, and the free Cu2+is easily reduced to Cu+by sodium ascorbate. Then Cu+catalyzes the cyclization reaction between the specially designed 5'-azide and 3'-alkyne tagged padlock probes through Cu+catalyzed azide-alkyne cycloaddition (CuAAC), which in turn initiates the hyperbranched rolling circle amplification (HRCA). Given that the CLT-HRCA products contain large amounts of double-stranded DNAs (dsDNAs), the addition of SYBR Green I resulted in the enhanced fluorescence signal. There was a linear relationship between the enhanced fluorescence intensity and the logarithm PPase activity ranging from 0.05 to 25 mU with a detection limit of 0.02 mU. Such proposed biosensor has been successfully applied to screen the potential PPase inhibitors and has accessed the related inhibit ability with high efficiency.
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
authors.forename: Yajuan
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China. lcuiying@fzu.edu.cn.
authors.forename: Cuiying
authors.initials: C
authors.lastname: Lin
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Lin
authors.affiliation: College of Biological Science and Technology, Fuzhou University, No. 2 Xueyuan Road, Fuzhou, Fujian, 350116, China. luofang0812@163.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Luo
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China.
authors.forename: Guonan
authors.initials: G
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques; D061565:Cycloaddition Reaction; D005456:Fluorescent Dyes / Q000737:chemistry*; D043564:Inorganic Pyrophosphatase / Q000032:analysis* / Q000037:antagonists & inhibitors; D057230:Limit of Detection; D013050:Spectrometry, Fluorescence
chemical_list: D005456:Fluorescent Dyes; D043564:Inorganic Pyrophosphatase
keywords: Click ligation trigger hyperbranched rolling circle amplification; Inorganic pyrophosphatase; Inorganic pyrophosphatase inhibitors