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Hydrogenase from Acetobacterium woodii

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Abstract

Hydrogenase from fructose-grown cells of Acetobacterium woodii has been purified 70-fold to a specific activity of 3,500 μmol hydrogen oxidized per min per mg of protein measured at 35°C and pH 7.6 with methyl viologen as electron acceptor. At the same conditions with reduced methyl viologen as electron donor the enzyme catalyzes the evolvement of 440 μmol of H2 per min per mg of protein. The enzyme was found in the soluble portion of the cell, indicating that it is either not membrane-bound or is loosely associated with the membrane. The purified enzyme, which does not contain nickel, exhibits spectroscopic properties similar to the iron-sulfur hydrogenase of Clostridium pasteurianum. The enzyme is strongly inhibited by carbon monoxide, with 50% inhibition occurring at approximately 7 nM CO. Ferredoxin, flavodoxin, and carbon monoxide dehydrogenase are reduced in hydrogen-dependent reaction by the A. woodi hydrogenase.

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Abbreviations

CO dehydrogenase:

carbon monoxide dehydrogenase

MV:

methyl viologen

SDS:

sodium dodecyl sulfate

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Author notes

  1. Steve W. Ragsdale

    Present address: Department of Biochemistry, Case Western Reserve University, 44106, Cleveland, OH, USA

Authors and Affiliations

  1. Department of Biochemistry, University of Georgia, 30602, Athens, GA, USA

    Steve W. Ragsdale & Lars G. Ljungdahl

Authors
  1. Steve W. Ragsdale
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  2. Lars G. Ljungdahl
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Additional information

This paper is dedicated to Professor Dr. Hans G. Schlegel on the occasion of his 60-years birthday. Hans' contributions to the field of microbiology are many and it is a pleasure for us to commemorate him in this way. One of us, L. G. L., had the fortune as a Humboldt-Preis recipient to spend a year at the Institut für Mikrobiologie der Universität Göttingen. Besides the best possible working conditions memories involve pleasant evenings with a glass of Rhine-wine in the Schlegels' home to strenuous back-packing in the Austrian Alps.

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Ragsdale, S.W., Ljungdahl, L.G. Hydrogenase from Acetobacterium woodii . Arch. Microbiol. 139, 361–365 (1984). https://doi.org/10.1007/BF00408380

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  • DOI: https://doi.org/10.1007/BF00408380

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