Structure and function of a second gene cluster encoding the formate dehydrogenase of Wolinella succinogenes

Abstract

Wolinella succinogenes contains a single formate dehydrogenase, but two gene loci (fdhI and fdhII)) code for the subunits of the enzyme. The nucleotide sequence of fdhII is almost identical with that of fdhI in the region comprising fdhEABCD. The sequences of fdhI and fdhII differ in the promotor regions upstream of fdhE. Deletion mutants lacking either fdhI or fdhII synthesize functional formate dehydrogenases, as shown by growth with formate as electron donor and either fumarate or polysulfide as acceptor substrates, and by the presence of the FdhA subunit and of enzyme activity. In the wild-type strain, the fdhI genes appear to be expressed preferentially during growth with formate and fumarate. The six-times greater amount of the enzyme present upon growth with formate and polysulfide is due to the expression of both fdhIand fdhII. The transcription start sites were located 196-bp and 129-bp upstream of the fdhE start codons of fdhI and fdhII, respectively. An apparently single transcript (5.6 kbp) was detected in polysulfide-grown W. succinogenes by Northern-blot analysis, suggesting that the five open reading frames form operons.