title: Amplified and selective detection of manganese peroxidase genes based on enzyme-scaffolded-gold nanoclusters and mesoporous carbon nitride.
abstract: This work has demonstrated an amplified and selective detection platform using enzyme-scaffolded-gold nanoclusters as signal label, coupling with mesoporous carbon nitride (MCN) and gold nanoparticles (GNPs) modified glassy carbon electrode (GCE). Streptavidin-horseradish peroxidase (SA-HRP) has been integrated with gold nanoclusters (GNCs) as scaffold using a simple, fast and non-toxic method. The mechanisms of enzymatic amplification, redox cycling and signal amplification by this biosensor were discussed in detail. GNCs might perform important roles as electrocatalyst as well as electron transducer in these processes. The concentrations of reagents and the reaction times of these reagents were optimized to improve the analytical performances. Under the optimized condition, the signal response to enzyme-scaffolded-gold nanoclusters catalyzed reaction was linearly related to the natural logarithm of the target nucleic acid concentration in the range from 10(-17)M to 10(-9)M with a correlation coefficient of 0.9946, and the detection limit was 8.0×10(-18)M (S/N=3). Besides, synthesized oligonucleotide as well as Phanerochaete chrysosporium MnP fragments amplified using polymerase chain reaction and digested by restriction endonucleases were tested. Furthermore, this biosensor exhibited good precision, stability, sensitivity, and selectivity, and discriminated satisfactorily against mismatched nucleic acid samples of similar lengths.
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Yaoyu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China. Electronic address: tanglin@hnu.edu.cn.
authors.forename: Lin
authors.initials: L
authors.lastname: Tang
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China. Electronic address: zgming@hnu.edu.cn.
authors.forename: Guangming
authors.initials: G
authors.lastname: Zeng
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Jiajia
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Changzheng
authors.initials: C
authors.lastname: Fan
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Guide
authors.initials: G
authors.lastname: Yang
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Environmental Science and Engineering, Hunan University, Changsha 410082, Hunan, PR China; Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, Hunan, PR China.
authors.forename: Xia
authors.initials: X
authors.lastname: Xie
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D017343:Genes, Plant; D006046:Gold / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D009570:Nitriles / Q000737:chemistry*; D010544:Peroxidases / Q000235:genetics*; D020075:Phanerochaete / Q000201:enzymology* / Q000235:genetics*; D016062:Porosity
chemical_list: D018744:DNA, Plant; D009570:Nitriles; C011206:cyanogen; D006046:Gold; D006735:Horseradish Peroxidase; D010544:Peroxidases; C051129:manganese peroxidase
keywords: Biosensor; Enzyme; Gold nanoclusters; Mesoporous carbon nitride; Signal amplification