Diffusion based kinetic selectivity modulation of enzymatic proteolysis in a microfluidic reactor : experimental analysis and stochastic modeling
| Autor: | Bertrand Bocquet, A. Treizebre, Renato Froidevaux, Adil Elagli, Simon Laurette |
|---|---|
| Přispěvatelé: | Laboratoire des Procédés Biologiques Génie Enzymatique et Microbien (PROBIOGEM), Université de Lille, Sciences et Technologies-Fédération Universitaire et Polytechnique de Lille, Institut d’Électronique, de Microélectronique et de Nanotechnologie - UMR 8520 (IEMN), Centrale Lille-Institut supérieur de l'électronique et du numérique (ISEN)-Université de Valenciennes et du Hainaut-Cambrésis (UVHC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Université Polytechnique Hauts-de-France (UPHF) |
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
0303 health sciences
education.field_of_study Chromatography Chemistry General Chemical Engineering Diffusion Microfluidics Kinetics Population Substrate (chemistry) Context (language use) Laminar flow 02 engineering and technology General Chemistry 021001 nanoscience & nanotechnology 03 medical and health sciences [SPI]Engineering Sciences [physics] Biophysics Microreactor 0210 nano-technology education 030304 developmental biology |
| Zdroj: | RSC Advances RSC Advances, 2014, 4, pp.3873-3882. ⟨10.1039/C3RA46005C⟩ RSC Advances, Royal Society of Chemistry, 2014, 4, pp.3873-3882. ⟨10.1039/C3RA46005C⟩ |
| ISSN: | 2046-2069 |
| Popis: | Microreactors enable new experimental ways for enzyme engineering. In this context, we show that the liquid–liquid parallel laminar flows in microchannels cause a kinetic selectivity modification of proteolytic enzymatic reaction involving hemoglobin and pepsin, reaction that generates numerous bioactive peptides at different advancement state. Here we show that this diffusion based kinetic modulation induces an altered peptides appearance kinetics for a part of the initial substrate population. Indeed, microfluidic and conventional batch experiments performed in the same reaction conditions lead to strong differences in the resulting peptidic profiles obtained by reversed-phase high-performance liquid chromatography. Such differences are explained by the laminar flow diffusive conditions inside microchannels and are supported by a stochastic algorithm based on the Michaelis–Menten equation. Several bioactive peptides are identified by mass spectrometry and show the potential of such methodology in peptides screening from a complex proteolysis but also for selective peptides preparation by microfluidics. |
| Databáze: | OpenAIRE |
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