Characterization and physiological roles of membrane-bound hydrogenase isoenzymes from Salmonella typhimurium.
We found that Salmonella typhimurium strain LT2 (Z) possessed two immunologically distinct, membrane-bound hydrogenase isoenzymes, which were similar in electrophoretic mobilities and apoprotein contents to hydrogenase isoenzymes 1 and 2 of Escherichia coli. The S. typhimurium enzymes cross-reacted with antibodies raised to the respective hydrogenase isoenzymes of E. coli. As for E. coli, an additional membrane-bound hydrogenase activity (termed hydrogenase 3), which did not cross-react with antibodies raised against either hydrogenase 1 or 2, was also present in detergent-dispersed membrane preparations. The physiological role of each of the three isoenzymes in E. coli has remained unclear owing to the lack of mutants specifically defective for individual isoenzymes. However, analysis of two additional wild-type isolates of S. typhimurium revealed specific defects in their hydrogenase isoenzyme contents. S. typhimurium LT2 (A) lacked isoenzyme 2 but possessed normal levels of hydrogenases 1 and 3. S. typhimurium LT7 lacked both isoenzymes 1 and 2 but retained normal hydrogenase 3 activity. Characterization of hydrogen metabolism by these hydrogenase-defective isolates allowed us to identify the physiological role of each of the three isoenzymes. Hydrogenase 3 activity correlated closely with formate hydrogenlyase-dependent hydrogen evolution, whereas isoenzyme 2 catalyzed hydrogen uptake (oxidation) during anaerobic, respiration-dependent growth. Isoenzyme 1 also functioned as an uptake hydrogenase but only during fermentative growth. We postulate that this enzyme functions in a hydrogen-recycling reaction which operates during fermentative growth.
D000693:Anaerobiosis; D002462:Cell Membrane / Q000201:enzymology; D005285:Fermentation; D005560:Formate Dehydrogenases / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000032:analysis / Q000378:metabolism*; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D009154:Mutation; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000254:growth & development
D005561:Formates; D007527:Isoenzymes; C030544:formic acid; D006859:Hydrogen; D006864:Hydrogenase; D005560:Formate Dehydrogenases; D005947:Glucose