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Rubredoxin from the green sulfur bacterium Chlorobaculum tepidum donates a redox equivalent to the flavodiiron protein in an NAD(P)H dependent manner via ferredoxin-NAD(P)+ oxidoreductase

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Abstract

The green sulfur bacterium, Chlorobaculum tepidum, is an anaerobic photoautotroph that performs anoxygenic photosynthesis. Although genes encoding rubredoxin (Rd) and a putative flavodiiron protein (FDP) were reported in the genome, a gene encoding putative NADH-Rd oxidoreductase is not identified. In this work, we expressed and purified the recombinant Rd and FDP and confirmed dioxygen reductase activity in the presence of ferredoxin-NAD(P)+ oxidoreductase (FNR). FNR from C. tepidum and Bacillus subtilis catalyzed the reduction of Rd at rates comparable to those reported for NADH-Rd oxidoreductases. Also, we observed substrate inhibition at high concentrations of NADPH similar to that observed with ferredoxins. In the presence of NADPH, B. subtilis FNR and Rd, FDP promoted dioxygen reduction at rates comparable to those reported for other bacterial FDPs. Taken together, our results suggest that Rd and FDP participate in the reduction of dioxygen in C. tepidum and that FNR can promote the reduction of Rd in this bacterium.

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Acknowledgements

The authors would like to thank ENAGO for the English language review.

Funding

This work was partly supported by Japan Society for the Promotion of Science KAKENHI Grant Number JP17K07304 (to DS) and JP18K06296 (to KI).

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Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, Kanagawa University, Tsuchiya, Hiratsuka, Kanagawa, 259-1293, Japan

    Wanwipa Ittarat, Takeshi Sato, Masaharu Kitashima & Kazuhito Inoue

  2. Research Institute for Integrated Science, Kanagawa University, Tsuchiya, Hiratsuka, Kanagawa, 259-1293, Japan

    Takeshi Sato, Hidehiro Sakurai & Kazuhito Inoue

  3. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Phahonyothin Road, Khlong Nueng, Khlong Luang, 12120, Pathum Thani, Thailand

    Wanwipa Ittarat

  4. Division of Material Science, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa, Ishikawa, 920-1192, Japan

    Daisuke Seo

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  1. Wanwipa Ittarat
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  2. Takeshi Sato
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  3. Masaharu Kitashima
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  4. Hidehiro Sakurai
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  5. Kazuhito Inoue
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Ittarat, W., Sato, T., Kitashima, M. et al. Rubredoxin from the green sulfur bacterium Chlorobaculum tepidum donates a redox equivalent to the flavodiiron protein in an NAD(P)H dependent manner via ferredoxin-NAD(P)+ oxidoreductase. Arch Microbiol 203, 799–808 (2021). https://doi.org/10.1007/s00203-020-02079-4

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