Chemical modifications by ionic liquid-inspired cations improve the activity and the stability of formate dehydrogenase in [MMIm][Me 2PO 4]

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][Me 2PO 4]) ionic liquid. In order to circumvent this problem, the enzyme wa...

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Veröffentlicht in:Journal of molecular catalysis. B, Enzymatic Enzymatic, 2010-08, Vol.65 (1), p.73-78
Hauptverfasser: Bekhouche, Mourad, Doumèche, Bastien, Blum, Loïc J.
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Sprache:eng
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Zusammenfassung: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][Me 2PO 4]) 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][Me 2PO 4] (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) ( t 1/2 of 9 days) and by a 3-fold factor after modification by ( 2) or ( 3) ( t 1/2 of 6 and 5 days respectively) in aqueous solution. When stored in 37.5% [MMIm][Me 2PO 4] (v:v), both modified and unmodified FDH have an increased half-life ( t 1/2 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.
ISSN:1381-1177
1873-3158
DOI:10.1016/j.molcatb.2010.01.028