Chemical modifications by ionic liquid-inspired cations improve the activity and the stability of formate dehydrogenase in [MMIm][Me2PO4] |
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| Affiliation: | 1. IFP Energies nouvelles, 1–4 Avenue de Bois Préau, 92852 Rueil-Malmaison, France;2. Institut des Sciences Chimiques de Rennes, UMR CNRS 6226, Université Rennes 1, 35042 Rennes, France;3. Institut Physique de Rennes, UMR CNRS 6251, Université Rennes 1, 35042 Rennes, France;4. Institut de Chimie de Clermont-Ferrand, ICCF, UMR CNRS 6296, BP 10448, F-63000 Clermont-Ferrand, France;1. Department of oral medicine and radiology, A. J. Institute of Dental Sciences, Kuntikan, Mangalore, 575004 Karnataka, India;2. Department of Pedodontia, A. J. Institute of Dental Sciences, Mangalore, India;3. Department of oral medicine and radiology, A. J. Institute of Dental Sciences, Mangalore, India;1. Energy Engineering, Division of Energy Science, Luleå University of Technology, 97187 Luleå, Sweden;2. Centre of Advanced Research in Bionanoconjugates and Biopolymers, Romanian Academy Petru Poni (PP) Institute of Macromolecular Chemistry, 00487 Iasi, Romania;3. Department of Materials and Environmental Chemistry, Arrhenius Laboratory, Stockholm University, SE-106 91 Stockholm, Sweden;4. State Key Laboratory of Materials-Oriented and Chemical Engineering, Nanjing Tech University, Nanjing 210009, PR China;5. Beijing Key Laboratory of Ionic Liquids Clean Process, CAS Key Laboratory of Green Process and Engineering, State Key Laboratory of Multiphase Complex Systems, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China;6. Dipartimento di Scienze Chimiche e Geologiche, Università degli Studi di Cagliari, 09042 Cagliari, Italy;1. Chemical and Materials Engineering, School of Engineering, University of Aberdeen, Aberdeen AB24 3UE, Scotland, UK;2. Chemical Engineering, School of Engineering & Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS, Scotland, UK;3. Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, Scotland, UK;4. School of Environmental Science and Engineering, Tianjin University, Tianjin 300072, China |
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| Abstract: | The formate dehydrogenase (FDH, EC: 1.2. 1.2) from Candida boidinii was found to be inactivated and unstable in the presence of high concentration (>50%) of the water soluble dimethylimidazolium dimethyl phosphate ([MMIm][Me2PO4]) ionic liquid. In order to circumvent this problem, the enzyme was chemically modified by cations usually present in ionic liquids: cholinium (1), hydroxyethyl-methylimidazolium (2) and hydroxypropyl-methylimidazolium (3) cations were activated with carbonyldiimidazole before being reacted with the FDH leading to a heterogeneous population of 6–7 biocatalysts. FDH modified by (1) or (3) led to 3–9 modifications while FDH modified by (2) led to 6 proteins presenting 7–12 grafted cations. Specific activity of the modified enzymes was decreased by a 2.5–3-fold factor (0.10–0.15 μmol min−1 mg−1) compared to the non-modified FDH (0.33 μmol min−1 mg−1) when assayed in carbonate buffer (pH 9.7, 25 mM). After modification, the FDH still present 0.06 μmol min−1 mg−1 in 70% [MMIm][Me2PO4] (v:v) (30–45% of their activity in aqueous buffer) while the native enzyme is inactive at this ionic liquid concentration, proving the efficiency of this strategy. The half-life of the modified enzyme is also increased by a 5-fold factor after modification by (1) (t1/2 of 9 days) and by a 3-fold factor after modification by (2) or (3) (t1/2 of 6 and 5 days respectively) in aqueous solution. When stored in 37.5% [MMIm][Me2PO4] (v:v), both modified and unmodified FDH have an increased half-life (t1/2 of 6–9 days). This grafting strategy is found to be good methods to mimic and study the stabilizing effect of ionic liquids on enzymes. |
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