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Développement d’un microréacteur biomimétique pour l'analyse in situ d'activités enzymatiques par couplage de l’électrochimie et de la microscopie de fluorescence

Abstract : Enzymatic reactions are involved in many physiological phenomena in living organisms. These reactions are based on protons and electrons transfers and can lead to the production of by-products. Among them, reactive oxygen and nitrogen species (ROS and RNS) are of great interest as they play a double role: on the one hand by allowing the organism to react to a stress by the activation of signaling redox pathways, and on the other hand, ROS and RNS can cause oxidative damages to tissues ensuing dysfunctions in the organism. The high reactivity of such species induce their short lifetimes (ns-min) and leads to uncertainties when it comes to the study of some enzymatic reactions in bulk. This PhD project aims to develop a biomimetic microreactor for the study of enzymatic ac-tivities producing ROS/RNS. Indeed, by confining a reaction within a cell-sized compartment (20-100 μm diameter), the generated species (H2O2, NO•, NO2-) could be analyzed in situ with a quantita-tive and kinetic resolution. Giant unilamellar vesicles are formed in physiological conditions and are used as microreactors for the monitoring of enzymatic activities of glucose oxidase and NO-synthases. Fluorescence microscopy allows individual vesicle observation and the monitoring of reactions trig-gered by microinjection. Then, released species are detected in real-time by electrochemistry in order to decipher the diverse enzymatic pathways of NO-Synthases.
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https://tel.archives-ouvertes.fr/tel-01680949
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Submitted on : Monday, January 22, 2018 - 9:49:07 AM
Last modification on : Tuesday, May 26, 2020 - 3:05:06 AM

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  • HAL Id : tel-01680949, version 2

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Pauline Lefrançois. Développement d’un microréacteur biomimétique pour l'analyse in situ d'activités enzymatiques par couplage de l’électrochimie et de la microscopie de fluorescence. Chimie théorique et/ou physique. Université de Bordeaux, 2017. Français. ⟨NNT : 2017BORD0759⟩. ⟨tel-01680949v2⟩

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