Stabilisation of the NAD⁺-reducing soluble [NiFe]-hydrogenase from Ralstonia eutropha H16 through modification with methoxy-poly(ethylene) glycol

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• 作者：Juliane  ; Ratzka^a ; Lars  ; Lauterbach^b ; Oliver  ; Lenz^b ; Marion B.  ; Ansorge-Schumacher^a ;   ; ^{m.ansorge@chem.tu-berlin.de}
• 关键词：NAD⁺ reducing hydrogenase ; Stability ; Organic solvents ; Ionic liquids ; mPEG modification
• 刊名：Journal of Molecular Catalysis B: Enzymatic
• 出版年：2012
• 出版时间：February 2012
• 年：2012
• 卷：74
• 期：3-4
• 页码：219-223
• 全文大小：377 K

文摘

It has lately been demonstrated that the NAD⁺-reducing soluble hydrogenase from Ralstonia eutropha H16 (SH) is a promising catalyst for the regeneration of NADH in biocatalysed asymmetric redox reactions. Such reactions often require the presence of water-miscible organic solvents and ionic liquids to enable efficient application to organic synthesis. In this study, we investigated the influence of frequently used solubilisers such as dimethyl sulphoxide [DMSO] and Tris (2-hydroxyethyl) methylammonium methylsulphate [MTEOA][MeSO₄] on the activity and stability of SH. The stability of the enzyme was significantly improved by covalent attachment of methoxy-poly(ethylene) glycol (mPEG). This modification led to significant increase of the half-life time from 0.1 to 0.5 h in the presence of 10 % (v/v) isopropanol. Interestingly, no stabilisation was observed for ionic liquids, while the activity of SH increased by up to 45.5 % . The mechanism(s) underlying these effects are discussed.