The function of Wolinella succinogenes psr genes in electron transport with polysulphide as the terminal electron acceptor

Abstract

The membrane-integrated polysulphide reductase (Psr) of Wolinella succinogenes is part of the electron transport chain catalyzing polysulphide reduction by formate or hydrogen. The isolated enzyme catalyzes sulphide oxidation by dimethylnaphthoquinone. The two hydrophilic subunits, PsrA and PsrB of the enzyme, are encoded by genes that form an apparent operon psrABC together with a third gene. Using homologous recombination, three deletion mutants of W. succinogenes were constructed that lack psrC, psrBC or the whole psr operon. The mutants grown with formate and fumarate were fractionated, and the cell fractions were analyzed for the presence of PsrA and enzyme activity. It was concluded that: (a) polysulphide reductase is a constituent of the wild-type chain catalyzing electron transport from formate to polysulphide; (b) the gene psrC encodes a subunit that anchors the enzyme in the membrane and is required for electron transport; (c) PsrA which probably carries the substrate site, is exposed to the bacterial periplasm; (d) PsrA and PsrB are required for the activity of sulphide oxidation with 2,3-dimethyl-1,4-naphthoquinone. Surprisingly, the delta psrABC mutant could grow with formate and polysulphide. The membrane fraction of the mutant grown under these conditions contained an enzyme that replaced polysulphide reductase in electron transport, and catalyzed sulphide oxidation with 2,3-dimethyl-1,4-naphthoquinone.