Differential expression of hydrogenase isoenzymes in Escherichia coli K-12: evidence for a third isoenzyme.
The cellular contents of the nickel-containing, membrane-bound hydrogenase isoenzymes 1 and 2 (hydrogenases 1 and 2) were analyzed by crossed immunoelectrophoresis. Their expression was differentially influenced by nutritional and genetic factors. Hydrogenase 2 content was enhanced after growth with either hydrogen and fumarate or glycerol and fumarate and correlated reasonably with cellular hydrogen uptake capacity. Hydrogenase 1 content was negligible under the above conditions but was enhanced by exogenous formate. Its expression was greatly reduced in a pfl mutant, which is unable to synthesise formate, but was restored to normal levels when the growth medium included formate. A mutation in the anaerobic regulatory gene, fnr, led to low overall hydrogenase activity and greatly reduced levels of both isoenzymes and abolished the formate enhancement of hydrogenase 1 content. Formate hydrogenlyase activity was similarly reduced in the fnr strain but, in contrast, was restored, as was overall hydrogenase activity, to normal levels by growth in the presence of formate. Low H2 uptake activity was found for the fnr strain under all growth conditions examined. Hydrogenase 1 content, therefore, does not correlate with formate hydrogenlyase activity and its role is unclear. A third hydrogenase isoenzyme, immunologically distinct from hydrogenases 1 and 2, whose expression is enhanced by formate, is present and forms part of the formate hydrogenlyase. We suggest that the effect of the fnr gene product on formate hydrogenlyase expression is mediated via internal formate.
D003429:Cross Reactions; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics*; D005561:Formates / Q000494:pharmacology; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005809:Genes, Regulator; D006864:Hydrogenase / Q000235:genetics*; D007123:Immunoelectrophoresis, Two-Dimensional; D007527:Isoenzymes / Q000235:genetics*; D009154:Mutation; D009532:Nickel
D029968:Escherichia coli Proteins; D005561:Formates; D007527:Isoenzymes; C030544:formic acid; D009532:Nickel; D006864:Hydrogenase; C000602631:hydrogenase 1, E coli