Engineered formate dehydrogenase from Chaetomium thermophilum , a promising enzymatic solution for biotechnical CO 2 fix
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ORIGINAL RESEARCH PAPER
Engineered formate dehydrogenase from Chaetomium thermophilum, a promising enzymatic solution for biotechnical CO2 fixation Mehmet M. C ¸ akar . Jouni Ruupunen . Juan Mangas-Sanchez . William R. Birmingham . Deniz Yildirim . Ossi Turunen . Nicholas J. Turner Jarkko Valjakka . Barıs¸ Binay
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Received: 10 December 2019 / Accepted: 5 June 2020 Ó Springer Nature B.V. 2020
Abstract Objectives Formate dehydrogenases (FDHs) are NAD(P)H-dependent enzymes that catalyse the reversible oxidation of formate to CO2. The main goal was to use directed evolution to obtain variants of the FDH from Chaetomium thermophilum (CtFDH) with enhanced reduction activity in the conversion of CO2 into formic acid. Results Four libraries were constructed targeting five residues in the active site. We identified two Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02937-7) contains supplementary material, which is available to authorized users.
variants (G93H/I94Y and R259C) with enhanced reduction activity which were characterised in the presence of both aqueous CO2(g) and HCO3-. The A1 variant (G93H/I94Y) showed a 5.4-fold increase in catalytic efficiency (kcat/KM) compared to that of the wild-type for HCO3- reduction. The improved biocatalysts were also applied as a coupled cofactor recycling system in the enantioselective oxidation of 4-phenyl-2-propanol catalysed by the alcohol dehydrogenase from Streptomyces coelicolor A3 (ScADH). Conversions in these reactions increased from 56 to 91% when the A1 variant was used instead of wildtype CtFDH.
M. M. C¸akar Department of Molecular Biology and Genetics, Gebze Technical University, 41400 Gebze, Kocaeli, Turkey
O. Turunen School of Forest Sciences, University of Eastern Finland, 80101 Joensuu, Finland
J. Ruupunen J. Valjakka Faculty of Medicine and Health Technology, Tampere University, 33014 Tampereen yliopisto, Finland
B. Binay (&) Department of Bioengineering, Gebze Technical University, 41400 Gebze, Kocaeli, Turkey e-mail: [email protected]
J. Mangas-Sanchez W. R. Birmingham N. J. Turner (&) Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK e-mail: [email protected] D. Yildirim Department of Chemistry, Cukurova University, 01330 Adana, Turkey
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Biotechnol Lett
Conclusions Two variants presenting up to five-fold increase in catalytic efficiency and kcat were obtained and characterised. They constitute a promising enzymatic alternative for CO2 utilization and will serve as scaffolds to be further developed in order to meet industrial requirements. Keywords Biotransformation CO2 reduction Enzyme engineering Molecular dynamics NAD?dependent FDH
Introduction Increasing human population, industrial activities and natural disasters (volcanic eruptions, etc.) have led to a continuous increase in the amount of carbon dioxide (CO2) released to the atmosph
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