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Pheromone synthesis in a biomicroreactor coated with anti-adsorption polyelectrolyte multilayer
Authors:Dimov Nikolay  Muñoz Lourdes  Carot-Sans Gerard  Verhoeven Michel L P M  Bula Wojciech P  Kocer Gülistan  Guerrero Angel  Gardeniers Han J G E
Institution:1Mesoscale Chemical Systems, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands;2Department of Biological Chemistry and Molecular Modeling (IQAC-CSIC), 08034 Barcelona, Spain
Abstract:To prepare a biosynthetic module in an infochemical communication project, we designed a silicon/glass microreactor with anti-adsorption polyelectrolyte multilayer coating and immobilized alcohol acetyl transferase (atf), one of the key biosynthetic enzymes of the pheromone of Spodoptera littoralis, on agarose beads inside. The system reproduces the last step of the biosynthesis in which the precursor diene alcohol (Z,E)-9,11-tetradecadienol is transformed into the major component (Z,E)-9,11-tetradecadienyl acetate. The scope of this study was to analyze and implement a multilayer, anti-adsorption coating based on layer-by-layer deposition of polyethylenimine/dextransulfate sodium salt (PEI/DSS). The multilayers were composed of two PEI with molecular weights 750 and 1.2 kDa at pH 9.2 or 6.0. Growth, morphology, and stability of the layers were analyzed by ellipsometry and atomic force microscopy (AFM). The anti-adsorption functionality of the multilayer inside the microreactor was validated. The activity of His(6)-(atf) was measured by gas chromatography coupled to mass spectrometer (GC-MS).
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