PMID: 40601Oct 11, 1979

Purification and properties of the particulate hydrogenase from the bacteroids of soybean root nodules

Biochimica Et Biophysica Acta
D J Arp, R H Burris

Abstract

The uptake hydrogenase (hydrogen:ferricytochrome c3 oxidoreductase, EC 1.12.2.1) from the bacteroids of soybean root nodules infected with Rhizobium japonicum 110 has been purified and characterized. Bacteroids were prepared, then broken by sonication. The particulate enzyme was solubilized by treatment with Triton X-100 and further purified by polyethylene glycol fractionation, DEAE-cellulose and Sephadex G-100 chromatography. The specific activity has been increased 196-fold to 19.6 units/mg protein. The molecular weight is 63 300 as determined by gel filtration and 65 300 as determined by SDS-polyacrylamide gel electrophoresis, indicating that the enzyme is a monomer. The enzyme is O2 sensitive, with a half-life of 70 min when exposed to air. The pH optimum of the solubilized enzyme is near 5.5; the Km for H2 is 1.4 microM. Suitable electron acceptors are methylene blue, ferricyanide, 2,6-dichlorophenolindophenol, and cytochrome c. Benzyl viologen is reduced slowly; methyl viologen, NAD(P)+, FAD, FMN, and O2 are not reduced. The optimum temperature for activity is 65-70 degrees C with an activation energy of 9.2 kcal. H2 evolution by the enzyme has been demonstrated. The hydrogenase is well-suited to function in an environme...Continue Reading

References

Jun 17, 1975·Biochemistry·P H Gitlitz, A I Krasna
Apr 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·K R Schubert, H J Evans

Citations

Oct 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M ZuberH J Evans
Oct 1, 1986·Plant Physiology·Y Kapulnik, D A Phillips
Oct 10, 2007·Biochemistry. Biokhimii︠a︡·A A TsygankovA A Karyakin
Jun 1, 1992·Analytical Biochemistry·D M Ferber, R J Maier
Mar 28, 1986·Biochemical and Biophysical Research Communications·P Boivin, C Bourdillon
Oct 17, 2007·FEMS Microbiology Ecology·Heather E Reed, Jennifer B H Martiny

Related Concepts

Cytochrome c Group
Energy Transfer
Hydrogen
Hydrogen-Ion Concentration
Oxidase
Rhizobium lupini
Glycine max

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