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Issue 1, 2019
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Hydrogel/enzyme dots as adaptable tool for non-compartmentalized multi-enzymatic reactions in microfluidic devices

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Abstract

This study presents a simple method to integrate hydrogel/enzyme dots for the performance of multi-enzymatic reactions in two microfluidic devices and shows the conversion of enzyme educts under continuous flow as well as the reusability of the hydrogel/enzyme dots in microfluidic devices. For this purpose, five different enzymes were physically entrapped in a hydrogel matrix composed of poly(ethylene glycol) diacrylate, 2-(dimethylamino)ethyl methacrylate, and 2-hydroxyethyl methacrylate. Two separate tri-enzymatic cascade reactions were carried out. In the first cascade the enzymes β-galactosidase, glucose oxidase, and horseradish peroxidase were used and the second cascade consisted of the enzymes phospholipase D, choline oxidase, and again horseradish peroxidase. The (long-term) activity of free and hydrogel-immobilized enzymes was evaluated by UV-vis spectroscopic measurements. Additionally, time-dependent enzyme leakage from the hydrogel was investigated. Following the successful execution of multi-enzymatic reactions in bulk hydrogels, the material was integrated into PDMS-on-glass microfluidic reactors to carry out the enzyme reactions in miniaturized scale and under continuous flow. For that, hydrogel dots with a diameter of 350 μm were covalently attached to planar glass substrates by UV-initiated polymerisation of the enzyme-containing hydrogel precursor. Experiments were carried out both in channel reactors with hydrogel dots arranged in rows and wide chamber reactors with a hexagonal array of hydrogels. Especially the latter one showed a good performance as the flow velocity and thus the shear force on the hydrogel was decreased. Additionally, the residence time of the substrate and consequently the yield were increased. Long-term activity of the tri-enzymatic reactions in microfluidic reactors was proven with an ABTS assay indicating that this approach may be used as a platform for the integration of (multi-)enzymatic hydrogel dot reactions in microfluidic systems without the need of additional modification steps.

Graphical abstract: Hydrogel/enzyme dots as adaptable tool for non-compartmentalized multi-enzymatic reactions in microfluidic devices

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Publication details

The article was received on 22 Aug 2018, accepted on 01 Nov 2018 and first published on 14 Nov 2018


Article type: Paper
DOI: 10.1039/C8RE00180D
Citation: React. Chem. Eng., 2019,4, 67-77
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    Hydrogel/enzyme dots as adaptable tool for non-compartmentalized multi-enzymatic reactions in microfluidic devices

    D. Simon, F. Obst, S. Haefner, T. Heroldt, M. Peiter, F. Simon, A. Richter, B. Voit and D. Appelhans, React. Chem. Eng., 2019, 4, 67
    DOI: 10.1039/C8RE00180D

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