Archives of Microbiology

, Volume 121, Issue 3, pp 199–206 | Cite as

Characteristics of the H2 oxidizing system in soybean nodule bacteroids

  • Tomás Ruiz-Argüeso
  • David W. Emerich
  • Harold J. Evans


A derivative of Rhizobium japonicum (strain 122 DES) has been isolated which forms nodules on soybeans that evolve little or no H2 in air and efficiently fixes N2. Bacteroids isolated from nodules formed by strain 122 DES took up H2 with O2 as the physiological acceptor and appeared to be typical of those R. japonicum strains that possess the H2 uptake system. The hydrogenase system in soybean nodules is located within the bacteroids and activity in macerated bacteroids is concentrated in a particulate fraction. The pH optimum for the reaction is near 8.0 and apparent Km values for H2 and O2 are 2 μM and 1 μM, respectively. The H2 oxidizing activity of a suspension of 122 DES bacteroids was stable at 4°C for at least 4 weeks and was not particularly sensitive to O2. Neither C2H2 nor CO inhibited O2 dependent H2 uptake activity.

Non-physiological electron acceptors of positive oxidation reduction potential also supported H2 uptake by bacteroids. The rate of H2 uptake with phenazine methosulfate as the acceptor was greater than that with O2. When methylene blue, triphenyltetrazolium, potassium ferricyanide or dichlorophenolindophenol were added to bacteriod suspensions, without preincubation, rates of H2 uptake were supported that were lower than those in the presence of O2. Preincubation of the bacteroids with acceptors increased the rates of H2 uptake. No H2 evolution was observed from reaction mixtures containing bacteroid suspensions and reduced methyl or benzyl viologens. Of a series of carbon substrates added to bacteroid suspensions only acetate, formate or succinate at concentrations of 50 mM resulted in 20% or greater inhibition of H2 oxidation.

The H2 uptake capacity of isolated 122 DES bacteroids (expressed on a dry bacteroid basis) was at least 10-fold higher than the rate of the nitrogenase reaction in nodules expressed on a comparable basis. Since about 1 mol of H2 is evolved for every mol of N2 reduced during the N2 fixation reaction, these observations explain why soybean nodules formed by strain 122 DES and other strains with high H2 uptake activities have a capacity for recycling all the H2 produced from the nitrogenase reaction.

Key words

Hydrogenase Hydrogen uptake Rhizobium japonicum Bacteroids Knallgas reaction 



PHenazine methosulfate


Methylene blue


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Copyright information

© Springer-Verlag 1979

Authors and Affiliations

  • Tomás Ruiz-Argüeso
    • 1
  • David W. Emerich
    • 2
  • Harold J. Evans
    • 2
  1. 1.Department of Microbiology, E.T.S. de Ingenieros AgrónomosUniversidad PolitécnicaMadridSpain
  2. 2.Laboratory for Nitrogen Fixation ResearchOregon State UniversityCorvallisUSA

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