Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Sep;131(3):922–928. doi: 10.1128/jb.131.3.922-928.1977

Hydrogenase Activity and the H2-Fumarate Electron Transport System in Bacteroides fragilis1

Martha A Harris 1, C Adinaryayana Reddy 1
PMCID: PMC235550  PMID: 893348

Abstract

Hydrogenase activity and the H2-fumarate electron transport system in a carbohydrate-fermenting obligate anaerobe, Bacteroides fragilis, were investigated. In both whole cells and cell extracts, hydrogenase activity was demonstrated with methylene blue, benzyl viologen, flavin mononucleotide, or flavin adenine dinucleotide as the electron acceptor. A catalytic quantity of benzyl viologen or ferredoxin from Clostridium pasteurianum was required to reduce nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate with H2. Much of the hydrogenase activity appeared to be associated with the soluble fraction of the cell. Fumarate reduction to succinate by H2 was demonstrable in cell extracts only in the presence of a catalytic quantity of benzyl viologen, flavin mononucleotide, flavin adenine dinucleotide, or ferredoxin from C. pasteurianum. Sulfhydryl compounds were not required for fumarate reduction by H2, but mercaptoethanol and dithiothreitol appeared to stimulate this activity by 59 and 61%, respectively. Inhibition of fumarate reduction by acriflavin, rotenone, 2-heptyl-4-hydroxyquinoline-N-oxide, and antimycin A suggest the involvement of a flavoprotein, a quinone, and cytochrome b in the reduction of fumarate to succinate. The involvement of a quinone in fumarate reduction is also apparent from the inhibition of fumarate reduction by H2 when cell extracts were irradiated with ultraviolet light. Based on the evidence obtained, a possible scheme for the flow of electrons from H2 to fumarate in B. fragilis is proposed.

Full text

PDF
922

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aspen A. J., Wolin M. J. Solubilization and reconstitution of a particulate hydrogenase from Vibrio succinogenes. J Biol Chem. 1966 Sep 25;241(18):4152–4156. [PubMed] [Google Scholar]
  2. BUCHANAN B. B., LOVENBERG W., RABINOWITZ J. C. A comparison of clostridial ferredoxins. Proc Natl Acad Sci U S A. 1963 Mar 15;49:345–353. doi: 10.1073/pnas.49.3.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barton L. L., Le Gall J., Peck H. D., Jr Phosphorylation coupled to oxidation of hydrogen with fumarate in extracts of the sulfate reducing bacterium, Desulfovibrio gigas. Biochem Biophys Res Commun. 1970 Nov 25;41(4):1036–1042. doi: 10.1016/0006-291x(70)90189-0. [DOI] [PubMed] [Google Scholar]
  4. Caldwell D. R., Bryant M. P. Medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria. Appl Microbiol. 1966 Sep;14(5):794–801. doi: 10.1128/am.14.5.794-801.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. GIBBONS R. J., ENGLE L. P. VITAMIN K COMPOUNDS IN BACTERIA THAT ARE OBLIGATE ANAEROBES. Science. 1964 Dec 4;146(3649):1307–1309. doi: 10.1126/science.146.3649.1307. [DOI] [PubMed] [Google Scholar]
  6. Hatefi Y. Flavoproteins of the electron transport system and the site of action of amytal, rotenone, and piericidin A. Proc Natl Acad Sci U S A. 1968 Jun;60(2):733–740. doi: 10.1073/pnas.60.2.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. JACOBS N. J., WOLIN M. J. Electron-transport system of Vibrio succinogenes. I. Enzymes and cytochromes of electron-transport system. Biochim Biophys Acta. 1963 Jan 1;69:18–28. doi: 10.1016/0006-3002(63)91221-6. [DOI] [PubMed] [Google Scholar]
  8. JACOBS N. J., WOLIN M. J. Electron-transport system of Vibrio succinogenes. II. Inhibition of electron transport by 2-heptyl-4-hydroxyquinoline N-oxide. Biochim Biophys Acta. 1963 Jan 1;69:29–39. doi: 10.1016/0006-3002(63)91222-8. [DOI] [PubMed] [Google Scholar]
  9. Kröger A., Dadák V., Klingenberg M., Diemer F. On the role of quinones in bacterial electron transport. Differential roles of ubiquinone and menaquinone in Proteus rettgeri. Eur J Biochem. 1971 Aug 16;21(3):322–333. doi: 10.1111/j.1432-1033.1971.tb01472.x. [DOI] [PubMed] [Google Scholar]
  10. Kröger A. Electron-transport phosphorylation coupled to fumarate reduction in anaerobically grown Proteus rettgeri. Biochim Biophys Acta. 1974 May 22;347(2):273–289. doi: 10.1016/0005-2728(74)90051-6. [DOI] [PubMed] [Google Scholar]
  11. LOESCHE W. J., SOCRANSKY S. S., GIBBONS R. J. BACTEROIDES ORALIS, PROPOSED NEW SPECIES ISOLATED FROM THE ORAL CAVITY OF MAN. J Bacteriol. 1964 Nov;88:1329–1337. doi: 10.1128/jb.88.5.1329-1337.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Macy J., Kulla H., Gottschalk G. H2-dependent anaerobic growth of Escherichia coli on L-malate: succinate formation. J Bacteriol. 1976 Feb;125(2):423–428. doi: 10.1128/jb.125.2.423-428.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moore W. E., Cato E. P., Holdeman L. V. Anaerobic bacteria of the gastrointestinal flora and their occurrence in clinical infections. J Infect Dis. 1969 Jun;119(6):641–649. doi: 10.1093/infdis/119.6.641. [DOI] [PubMed] [Google Scholar]
  15. PECK H. D., Jr, GEST H. Hydrogenase of Clostridium butylicum. J Bacteriol. 1957 Apr;73(4):569–580. doi: 10.1128/jb.73.4.569-580.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prasad R., Kalra V. K., Brodie A. F. Different mechanisms of energy coupling for transport of various amino acids in cells of Mycobacterium phlei. J Biol Chem. 1976 Apr 25;251(8):2493–2498. [PubMed] [Google Scholar]
  17. Reddy C. A., Bryant M. P., Wolin M. J. Ferredoxin-dependent conversion of acetaldehyde to acetate and H 2 in extracts of S organism. J Bacteriol. 1972 Apr;110(1):133–138. doi: 10.1128/jb.110.1.133-138.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tzeng S. F., Wolfe R. S., Bryant M. P. Factor 420-dependent pyridine nucleotide-linked hydrogenase system of Methanobacterium ruminantium. J Bacteriol. 1975 Jan;121(1):184–191. doi: 10.1128/jb.121.1.184-191.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. WOLIN M. J., WOLIN E. A., JACOBS N. J. Cytochrome-producing anaerobic Vibrio succinogenes, sp. n. J Bacteriol. 1961 Jun;81:911–917. doi: 10.1128/jb.81.6.911-917.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. White D. C., Sinclair P. R. Branched electron-transport systems in bacteria. Adv Microb Physiol. 1971;5:173–211. doi: 10.1016/s0065-2911(08)60407-5. [DOI] [PubMed] [Google Scholar]
  21. Yoch D. C., Valentine R. C. Ferredoxins and flavodoxins of bacteria. Annu Rev Microbiol. 1972;26:139–162. doi: 10.1146/annurev.mi.26.100172.001035. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES