Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans
Fixation of CO2 into the organic compound formate by formate dehydrogenases (FDHs) is regarded as the oldest autotrophic process on Earth. It has been proposed that an FDH-dependent CO2 fixation module could support CO2 assimilation even in photoautotrophic organisms. In the present study, we charac...
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Frontiers Media S.A.
2025-01-01
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| author | Eva-Maria Brouwer Hitesh K. R. Medipally Hitesh K. R. Medipally Saskia Schwab Shanshan Song Marc M. Nowaczyk Marc M. Nowaczyk Marc M. Nowaczyk Martin Hagemann Martin Hagemann |
| author_facet | Eva-Maria Brouwer Hitesh K. R. Medipally Hitesh K. R. Medipally Saskia Schwab Shanshan Song Marc M. Nowaczyk Marc M. Nowaczyk Marc M. Nowaczyk Martin Hagemann Martin Hagemann |
| author_sort | Eva-Maria Brouwer |
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| description | Fixation of CO2 into the organic compound formate by formate dehydrogenases (FDHs) is regarded as the oldest autotrophic process on Earth. It has been proposed that an FDH-dependent CO2 fixation module could support CO2 assimilation even in photoautotrophic organisms. In the present study, we characterized FDH from Clostridium carboxidivorans (ccFDH) due to its ability to reduce CO2 under aerobic conditions. During the production of recombinant ccFDH, in which the selenocysteine codon was replaced by Cys, we were able to replace the W with Mo as the transition metal in the ccFDH metal cofactor, resulting in a two-fold increase of 6 μmol formate min−1 in enzyme activity. Then, we generated ccFDH variants in which the strict NADH preference of the enzyme was changed to NADPH, as this reducing agent is produced in high amounts during the photosynthetic light process. Finally, we showed that the native ccFDH can also directly use ferredoxin as a reducing agent, which is produced by the photosynthetic light reactions at photosystem I. These data collectively suggest that ccFDH and, particularly, its optimized variants can be regarded as suitable enzymes to couple formate production to photosynthesis in photoautotroph organisms, which could potentially support CO2 assimilation via the Calvin–Benson–Bassham (CBB) cycle and minimize CO2 losses due to photorespiration. |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj-art-8b481e33c70e4a45baa354fc8e387a9a2025-01-13T06:10:47ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011510.3389/fmicb.2024.15276261527626Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivoransEva-Maria Brouwer0Hitesh K. R. Medipally1Hitesh K. R. Medipally2Saskia Schwab3Shanshan Song4Marc M. Nowaczyk5Marc M. Nowaczyk6Marc M. Nowaczyk7Martin Hagemann8Martin Hagemann9Department of Plant Physiology, Institute of Biosciences, University of Rostock, Rostock, GermanyDepartment of Plant Biochemistry, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Bochum, GermanyScience for Life Laboratory, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH—Royal Institute of Technology, Stockholm, SwedenDepartment of Plant Physiology, Institute of Biosciences, University of Rostock, Rostock, GermanyDepartment of Plant Physiology, Institute of Biosciences, University of Rostock, Rostock, GermanyDepartment of Plant Biochemistry, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Bochum, GermanyDepartment of Biochemistry, Institute of Biosciences, University of Rostock, Rostock, GermanyDepartment of Life, Light and Matter, Interdisciplinary Faculty, University of Rostock, Rostock, GermanyDepartment of Plant Physiology, Institute of Biosciences, University of Rostock, Rostock, GermanyDepartment of Life, Light and Matter, Interdisciplinary Faculty, University of Rostock, Rostock, GermanyFixation of CO2 into the organic compound formate by formate dehydrogenases (FDHs) is regarded as the oldest autotrophic process on Earth. It has been proposed that an FDH-dependent CO2 fixation module could support CO2 assimilation even in photoautotrophic organisms. In the present study, we characterized FDH from Clostridium carboxidivorans (ccFDH) due to its ability to reduce CO2 under aerobic conditions. During the production of recombinant ccFDH, in which the selenocysteine codon was replaced by Cys, we were able to replace the W with Mo as the transition metal in the ccFDH metal cofactor, resulting in a two-fold increase of 6 μmol formate min−1 in enzyme activity. Then, we generated ccFDH variants in which the strict NADH preference of the enzyme was changed to NADPH, as this reducing agent is produced in high amounts during the photosynthetic light process. Finally, we showed that the native ccFDH can also directly use ferredoxin as a reducing agent, which is produced by the photosynthetic light reactions at photosystem I. These data collectively suggest that ccFDH and, particularly, its optimized variants can be regarded as suitable enzymes to couple formate production to photosynthesis in photoautotroph organisms, which could potentially support CO2 assimilation via the Calvin–Benson–Bassham (CBB) cycle and minimize CO2 losses due to photorespiration.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1527626/fullenzymephotosynthesisredoxsite-specific mutantscarbon fixation |
| spellingShingle | Eva-Maria Brouwer Hitesh K. R. Medipally Hitesh K. R. Medipally Saskia Schwab Shanshan Song Marc M. Nowaczyk Marc M. Nowaczyk Marc M. Nowaczyk Martin Hagemann Martin Hagemann Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans Frontiers in Microbiology enzyme photosynthesis redox site-specific mutants carbon fixation |
| title | Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans |
| title_full | Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans |
| title_fullStr | Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans |
| title_full_unstemmed | Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans |
| title_short | Characterization of the oxygen-tolerant formate dehydrogenase from Clostridium carboxidivorans |
| title_sort | characterization of the oxygen tolerant formate dehydrogenase from clostridium carboxidivorans |
| topic | enzyme photosynthesis redox site-specific mutants carbon fixation |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1527626/full |
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