Carbon Fixation in the Chemolithoautotrophic Bacterium Aquifex aeolicus Involves Two Low-Potential Ferredoxins as Partners of the PFOR and OGOR Enzymes

Laura Prioretti (1) , Giulia d'Ermo (1) , Pascale Infossi (1) , Arlette Kpebe (1) , Régine Lebrun (2) , Marielle Bauzan (2) , Elisabeth Lojou (1) , Bruno Guigliarelli (1) , Marie-Thérèse Giudici-Orticoni (1) , Marianne Guiral (1)

1 BIP - Bioénergétique et Ingénierie des Protéines
2 IMM - Institut de Microbiologie de la Méditerranée

Aquifex aeolicus is a microaerophilic hydrogen-and sulfur-oxidizing bacterium that assimilates CO 2 via the reverse tricarboxylic acid cycle (rTCA). Key enzymes of this pathway are pyruvate:ferredoxin oxidoreductase (PFOR) and 2-oxoglutarate:ferredoxin oxidoreductase (OGOR), which are responsible, respectively, for the reductive carboxylation of acetyl-CoA to pyruvate and of succinyl-CoA to 2-oxoglutarate, two energetically unfavorable reactions that require a strong reduction potential. We have confirmed, by biochemistry and proteomics, that A. aeolicus possesses a pentameric version of these enzyme complexes ((αβγδε) 2) and that they are highly abundant in the cell. In addition, we have purified and characterized, from the soluble fraction of A. aeolicus, two low redox potential and oxygen-stable [4Fe-4S] ferredoxins (Fd6 and Fd7, E 0 = −440 and −460 mV, respectively) and shown that they can physically interact and exchange electrons with both PFOR and OGOR, suggesting that they could be the physiological electron donors of the system in vivo. Shotgun proteomics indicated that all the enzymes assumed to be involved in the rTCA cycle are produced in the A. aeolicus cells. A number of additional enzymes, previously suggested to be part of a putative partial Wood-Ljungdahl pathway used for the synthesis of serine and glycine from CO 2 were identified by mass spectrometry, but their abundance in the cell seems to be much lower than that of the rTCA cycle. Their possible involvement in carbon assimilation is discussed.

Mots clés: Wood-Ljungdahl pathway; carbon fixation; chemolithoautotrophic bacteria; hydrogenase; hyperthermophilic bacteria; low potential ferredoxin; oxoglutarate:ferredoxin oxidoreductase; pyruvate:ferredoxin oxidoreductase; reverse TCA cycle.

Domaines: Bactériologie; Biochimie [q-bio.BM]

Laura Prioretti, Giulia d'Ermo, Pascale Infossi, Arlette Kpebe, Régine Lebrun, et al.. Carbon Fixation in the Chemolithoautotrophic Bacterium Aquifex aeolicus Involves Two Low-Potential Ferredoxins as Partners of the PFOR and OGOR Enzymes. Life, 2023, 13 (3), pp.627. ⟨10.3390/life13030627⟩. ⟨hal-04071170⟩