respiratory membrane endo-hydrogenase activity in the microaerophile azorhizobium caulinodans is bidirectional呼吸膜endo-hydrogenase活动microaerophile azorhizobium caulinodans是双向的.pdfVIP

Respiratory Membrane endo-Hydrogenase Activity in the Microaerophile Azorhizobium caulinodans Is Bidirectional Brittany N. Sprecher, Margo E. Gittings, Robert A. Ludwig* Sinsheimer Laboratories, Department of Molecular, Cellular and Developmental Biology, University of California Santa Cruz, Santa Cruz, California, United States of America Abstract Background: The microaerophilic bacterium Azorhizobium caulinodans, when fixing N2 both in pure cultures held at 20 mM dissolved O2 tension and as endosymbiont of Sesbania rostrata legume nodules, employs a novel, respiratory-membrane endo-hydrogenase to oxidize and recycle endogenous H2 produced by soluble Mo-dinitrogenase activity at the expense of O2. Methods and Findings: From a bioinformatic analysis, this endo-hydrogenase is a core (6 subunit) version of (14 subunit) NADH:ubiquinone oxidoreductase (respiratory complex I). In pure A. caulinodans liquid cultures, when O2 levels are lowered to ,1 mM dissolved O2 tension (true microaerobic physiology), in vivo endo-hydrogenase activity reverses and continuously evolves H2 at high rates. In essence, H+ ions then supplement scarce O2 as respiratory-membrane electron acceptor. Paradoxically, from thermodynamic considerations, such hydrogenic respiratory-membrane electron transfer need largely uncouple oxidative phosphorylation, required for growth of non-phototrophic aerobic bacteria, A. caulinodans included. Conclusions: A. caulinodans in vivo endo-hydrogenase catalytic activity is bidirectional. To our knowledge, this study is the first demonstration of hydrogenic respiratory-membrane electron transfer among aerobic (non-fermentative) bacteria. When compared with O2 tolerant hydrogenases in other organisms, A. cau