Organohalide respiration in Dehalococcoides strains represents a novel mode of proton motive force generation.

Abstract

Dehalococcoides strains grow obligately by respiration with hydrogen as an electron donor and halogenated compounds as terminal electron acceptors, catalysed by a single membrane-integrated protein supercomplex. Many insights have been gained into the respiratory complex based on physiological experiments, biochemical analyses, genome sequencing, and proteomics. Recent data acquired from activity tests with deuterated water and whole cells revealed the mode of energy conservation by this respiratory complex. The data shows that the proton required for periplasmic dehalogenation originates from inside the cell, suggesting an electrogenic protonation of the electron acceptor, while two protons are released into the periplasm by hydrogen oxidation. This surprisingly simple mechanism of pmf generation aligns with the subunit composition of the respiratory complex, the orientation of the subunits in the membrane, the absence of quinones as electron mediators, the rigidity of the cell membrane, as evidenced by its phospholipid fatty acid composition, and with proton channels formed by protonatable amino acid residues identified in the AlphaFold2-predicted structure of one of the membrane-spanning subunits. The respiration model is characterised by: (i) electrogenic protonation of the electron acceptor; (ii) reliance on a single protein complex for pmf generation without quinones; (iii) lack of transmembrane cytochromes; (iv) presence of both redox-active centres on the same side of the membrane, both facing the periplasm; and (v) restriction of the electron flow to periplasmic subunits of the respiratory complex. This type of respiration may represent an ancestral, quinone-free mechanism, offering inspiring new biotechnological applications.