title: Sensitive biosensor based on recombinant PP1α for microcystin detection.
abstract: A novel electrochemical microcystin-LR (MC-LR) biosensor based on the inhibition of recombinant protein phosphate type 1 (PP1α) is reported in this work. The use of innovative recombinant enzyme led to investigate new commercially available substrate, electrochemically active after their dephosphorylation by PP1α. Only two of selected substrates, 1-naphylphosphate and phosphoparacetamol, showed a good affinity toward PP1α. Kinetic parameters were performed by classical colorimetric assays and revealed that phosphoparacetamol is an excellent synthetic substrate with a Km value of 1.2 mM. The reported biosensor is constructed by entrapment of the enzyme in Polyvinyl Alcohol (azid unit) on Cobalt-Phtalocyanine (CoPC) modified screen printed electrode. Electrocatalytic mediator demonstrated a significant improvement in the electrochemical detection of dephosphorylated substrate. The standard inhibition curve has provided a limit of detection at 0.93 µg/L and a broad dynamic range from 0.93 to 40.32 μg/L for MC-LR, demonstrating the improved analytical performance.
authors.affiliation: Laboratoire IMAgES (Institut de Modélisation et d'Analyses en Géo-Environnements et Santé (EA 4218)) - Université de Perpignan Via Domitia, Bat S, 52 avenue Paul alduy, 66860 Perpignan Cedex, France. Electronic address: gaelle.catanante@univ-perp.fr.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Catanante
authors.affiliation: Laboratoire IMAgES (Institut de Modélisation et d'Analyses en Géo-Environnements et Santé (EA 4218)) - Université de Perpignan Via Domitia, Bat S, 52 avenue Paul alduy, 66860 Perpignan Cedex, France. Electronic address: laura.espin@hotmail.fr.
authors.forename: Laura
authors.initials: L
authors.lastname: Espin
authors.affiliation: Laboratoire IMAgES (Institut de Modélisation et d'Analyses en Géo-Environnements et Santé (EA 4218)) - Université de Perpignan Via Domitia, Bat S, 52 avenue Paul alduy, 66860 Perpignan Cedex, France. Electronic address: jlmarty@univ-perp.fr.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Marty
mesh_terms: D015374:Biosensing Techniques; D003124:Colorimetry; D004566:Electrodes; D052998:Microcystins / Q000737:chemistry / Q000302:isolation & purification*; D054645:Protein Phosphatase 1 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D052998:Microcystins; D011994:Recombinant Proteins; C078588:microcystin; D054645:Protein Phosphatase 1
keywords: Biosensor; Electrochemical detection; Environmental analysis; Microcystins; Recombinant protein phosphatase (PP1α)