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Engineered formate dehydrogenase from Chaetomium thermophilum, a promising enzymatic solution for biotechnical CO2 fixation.

Authors
  • Çakar, Mehmet M1
  • Ruupunen, Jouni2
  • Mangas-Sanchez, Juan3
  • Birmingham, William R3
  • Yildirim, Deniz4
  • Turunen, Ossi5
  • Turner, Nicholas J6
  • Valjakka, Jarkko2
  • Binay, Barış7
  • 1 Department of Molecular Biology and Genetics, Gebze Technical University, 41400, Gebze, Kocaeli, Turkey. , (Turkey)
  • 2 Faculty of Medicine and Health Technology, Tampere University, 33014, Tampereen yliopisto, Finland. , (Finland)
  • 3 Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
  • 4 Department of Chemistry, Cukurova University, 01330, Adana, Turkey. , (Turkey)
  • 5 School of Forest Sciences, University of Eastern Finland, 80101, Joensuu, Finland. , (Finland)
  • 6 Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK. [email protected]
  • 7 Department of Bioengineering, Gebze Technical University, 41400, Gebze, Kocaeli, Turkey. [email protected] , (Turkey)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Jun 16, 2020
Identifiers
DOI: 10.1007/s10529-020-02937-7
PMID: 32557118
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

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. Four libraries were constructed targeting five residues in the active site. We identified two 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 wild-type CtFDH. 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.

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