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Carboxylation reaction catalyzed by 2-oxoglutarate:ferredoxin oxidoreductases from Hydrogenobacter thermophilus

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Abstract

Hydrogenobacter thermophilus TK-6 is a thermophilic, chemolithoautotrophic, hydrogen-oxidizing bacterium that fixes carbon dioxide via the reductive tricarboxylic acid (rTCA) cycle. 2-Oxoglutarate:ferredoxin oxidoreductase (OGOR) is the key enzyme in this cycle that fixes carbon dioxide. The genome of strain TK-6 encodes at least two distinct OGOR enzymes, termed For and Kor. We report here a method for measuring the carboxylation of succinyl-CoA catalyzed by OGORs. The method involves the in vitro coupling of OGOR with ferredoxin and pyruvate:ferredoxin oxidoreductase from strain TK-6, and glutamate dehydrogenase from Sulfolobus tokodaii. Using this method, we determined both the apparent maximum velocities and the Km values of For and Kor for the carboxylation of succinyl-CoA. This is the first reported kinetic analysis of carbon fixation catalyzed by OGOR enzymes from the rTCA cycle.

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Abbreviations

rTCA:

Reductive tricarboxylic acid

OGOR:

2-Oxoglutarate:ferredoxin oxidoreductase

OR:

2-Oxoacid oxidoreductase

POR:

Pyruvate:ferredoxin oxidoreductase

GDH:

Glutamate dehydrogenase

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Authors and Affiliations

  1. Department of Biotechnology, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo, 113-8657, Japan

    Masahiro Yamamoto, Takeshi Ikeda, Hiroyuki Arai, Masaharu Ishii & Yasuo Igarashi

Authors
  1. Masahiro Yamamoto
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  2. Takeshi Ikeda
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  3. Hiroyuki Arai
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  4. Masaharu Ishii
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  5. Yasuo Igarashi
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Corresponding author

Correspondence to Masaharu Ishii.

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Communicated by F. Robb.

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Yamamoto, M., Ikeda, T., Arai, H. et al. Carboxylation reaction catalyzed by 2-oxoglutarate:ferredoxin oxidoreductases from Hydrogenobacter thermophilus . Extremophiles 14, 79–85 (2010). https://doi.org/10.1007/s00792-009-0289-4

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  • DOI: https://doi.org/10.1007/s00792-009-0289-4

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