A purified energy-converting hydrogenase from Thermoanaerobacter kivui demonstrates coupled H⁺-translocation and reduction in vitro.

ABSTRACT: Energy-converting hydrogenases (Ech) are ancient, membrane-bound enzymes that use reduced ferredoxin (Fd) as an electron donor to reduce protons to molecular H₂. Experiments with whole cells, membranes and vesicle-fractions suggest that proton reduction is coupled to proton translocation across the cytoplasmatic membrane, but this has never been demonstrated with a purified enzyme. To this end, we produced a His-tagged Ech complex in the thermophilic and anaerobic bacterium Thermoanaerobacter kivui. The enzyme could be purified by affinity chromatography from solubilized membranes with full retention of its eight subunits, as well as full retention of physiological activities, i.e., H₂-dependent Fd reduction and Fd^2--dependent H₂ production. We f

SUBMITTER: Katsyv A 

PROVIDER: S-EPMC9356269 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature