Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Jul;171(7):3854–3859. doi: 10.1128/jb.171.7.3854-3859.1989

Reconstitution and properties of a coenzyme F420-mediated formate hydrogenlyase system in Methanobacterium formicicum.

S F Baron 1, J G Ferry 1
PMCID: PMC210135  PMID: 2661536

Abstract

Formate hydrogenlyase activity in a cell extract of Methanobacterium formicicum was abolished by removal of coenzyme F420; addition of purified coenzyme F420 restored activity. Formate hydrogenlyase activity was reconstituted with three purified components from M. formicicum: coenzyme F420-reducing hydrogenase, coenzyme F420-reducing formate dehydrogenase, and coenzyme F420. The reconstituted system required added flavin adenine dinucleotide (FAD) for maximal activity. Without FAD, the formate dehydrogenase and hydrogenase rapidly lost coenzyme F420-dependent activity relative to methyl viologen-dependent activity. Immunoadsorption of formate dehydrogenase or coenzyme F420-reducing hydrogenase from the cell extract greatly reduced formate hydrogenlyase activity; addition of the purified enzymes restored activity. The formate hydrogenlyase activity was reversible, since both the cell extract and the reconstituted system produced formate from H2 plus CO2 and HCO3-.

Full text

PDF

Selected References

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

  1. Balch W. E., Fox G. E., Magrum L. J., Woese C. R., Wolfe R. S. Methanogens: reevaluation of a unique biological group. Microbiol Rev. 1979 Jun;43(2):260–296. doi: 10.1128/mr.43.2.260-296.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barber M. J., Siegel L. M., Schauer N. L., May H. D., Ferry J. G. Formate dehydrogenase from Methanobacterium formicicum. Electron paramagnetic resonance spectroscopy of the molybdenum and iron-sulfur centers. J Biol Chem. 1983 Sep 25;258(18):10839–10845. [PubMed] [Google Scholar]
  3. Baron S. F., Brown D. P., Ferry J. G. Locations of the hydrogenases of Methanobacterium formicicum after subcellular fractionation of cell extract. J Bacteriol. 1987 Aug;169(8):3823–3825. doi: 10.1128/jb.169.8.3823-3825.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baron S. F., Ferry J. G. Purification and properties of the membrane-associated coenzyme F420-reducing hydrogenase from Methanobacterium formicicum. J Bacteriol. 1989 Jul;171(7):3846–3853. doi: 10.1128/jb.171.7.3846-3853.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  6. Ferry J. G., Wolfe R. S. Nutritional and biochemical characterization of Methanospirillum hungatii. Appl Environ Microbiol. 1977 Oct;34(4):371–376. doi: 10.1128/aem.34.4.371-376.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fox J. A., Livingston D. J., Orme-Johnson W. H., Walsh C. T. 8-Hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum: 1. Purification and characterization. Biochemistry. 1987 Jul 14;26(14):4219–4227. doi: 10.1021/bi00388a007. [DOI] [PubMed] [Google Scholar]
  8. GRAY C. T., GEST H. BIOLOGICAL FORMATION OF MOLECULAR HYDROGEN. Science. 1965 Apr 9;148(3667):186–192. doi: 10.1126/science.148.3667.186. [DOI] [PubMed] [Google Scholar]
  9. Jacobson F. S., Daniels L., Fox J. A., Walsh C. T., Orme-Johnson W. H. Purification and properties of an 8-hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum. J Biol Chem. 1982 Apr 10;257(7):3385–3388. [PubMed] [Google Scholar]
  10. Jones J. B., Stadtman T. C. Reconstitution of a formate-NADP+ oxidoreductase from formate dehydrogenase and a 5-deazaflavin-linked NADP+ reductase isolated from Methanococcus vannielii. J Biol Chem. 1980 Feb 10;255(3):1049–1053. [PubMed] [Google Scholar]
  11. Lespinat P. A., Berlier Y., Fauque G., Czechowski M., Dimon B., Le Gall J. The pH dependence of proton-deuterium exchange, hydrogen production and uptake catalyzed by hydrogenases from sulfate-reducing bacteria. Biochimie. 1986 Jan;68(1):55–61. doi: 10.1016/s0300-9084(86)81068-9. [DOI] [PubMed] [Google Scholar]
  12. Lovley D. R., Greening R. C., Ferry J. G. Rapidly growing rumen methanogenic organism that synthesizes coenzyme M and has a high affinity for formate. Appl Environ Microbiol. 1984 Jul;48(1):81–87. doi: 10.1128/aem.48.1.81-87.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. May H. D., Patel P. S., Ferry J. G. Effect of molybdenum and tungsten on synthesis and composition of formate dehydrogenase in Methanobacterium formicicum. J Bacteriol. 1988 Aug;170(8):3384–3389. doi: 10.1128/jb.170.8.3384-3389.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. May H. D., Schauer N. L., Ferry J. G. Molybdopterin cofactor from Methanobacterium formicicum formate dehydrogenase. J Bacteriol. 1986 May;166(2):500–504. doi: 10.1128/jb.166.2.500-504.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Muth E., Mörschel E., Klein A. Purification and characterization of an 8-hydroxy-5-deazaflavin-reducing hydrogenase from the archaebacterium Methanococcus voltae. Eur J Biochem. 1987 Dec 15;169(3):571–577. doi: 10.1111/j.1432-1033.1987.tb13647.x. [DOI] [PubMed] [Google Scholar]
  16. Nagle D. P., Jr, Wolfe R. S. Component A of the methyl coenzyme M methylreductase system of Methanobacterium: resolution into four components. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2151–2155. doi: 10.1073/pnas.80.8.2151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nelson M. J., Brown D. P., Ferry J. G. FAD requirement for the reduction of coenzyme F420 by hydrogenase from Methanobacterium formicicum. Biochem Biophys Res Commun. 1984 May 16;120(3):775–781. doi: 10.1016/s0006-291x(84)80174-6. [DOI] [PubMed] [Google Scholar]
  18. PECK H. D., Jr, GEST H. Formic dehydrogenase and the hydrogenlyase enzyme complex in coli-aerogenes bacteria. J Bacteriol. 1957 Jun;73(6):706–721. doi: 10.1128/jb.73.6.706-721.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Patel P. S., Ferry J. G. Characterization of the upstream region of the formate dehydrogenase operon of Methanobacterium formicicum. J Bacteriol. 1988 Aug;170(8):3390–3395. doi: 10.1128/jb.170.8.3390-3395.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Petrov R. R., Utkin I. B., Popov V. O. Redox-dependent inactivation of the NAD-dependent hydrogenase from Alcaligenes eutrophus Z1. Arch Biochem Biophys. 1989 Jan;268(1):298–305. doi: 10.1016/0003-9861(89)90591-2. [DOI] [PubMed] [Google Scholar]
  21. Qadri S. M., Hoare D. S. Formic hydrogenlyase and the photoassimilation of formate by a strain of Rhodopseudomonas palustris. J Bacteriol. 1968 Jun;95(6):2344–2357. doi: 10.1128/jb.95.6.2344-2357.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reddy C. A., Bryant M. P., Wolin M. J. Ferredoxin- and nicotinamide adenine dinucleotide-dependent H 2 production from ethanol and formate in extracts of S organism isolated from "Methanobacillus omelianskii". J Bacteriol. 1972 Apr;110(1):126–132. doi: 10.1128/jb.110.1.126-132.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. STADTMAN T. C., BARKER H. A. Studies on the methane fermentation. X. A new formate-decomposing bacterium, Methanococcus vannielii. J Bacteriol. 1951 Sep;62(3):269–280. doi: 10.1128/jb.62.3.269-280.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schauer N. L., Ferry J. G. Composition of the coenzyme F420-dependent formate dehydrogenase from Methanobacterium formicicum. J Bacteriol. 1986 Feb;165(2):405–411. doi: 10.1128/jb.165.2.405-411.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schauer N. L., Ferry J. G. FAD requirement for the reduction of coenzyme F420 by formate dehydrogenase from Methanobacterium formicicum. J Bacteriol. 1983 Aug;155(2):467–472. doi: 10.1128/jb.155.2.467-472.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schauer N. L., Ferry J. G., Honek J. F., Orme-Johnson W. H., Walsh C. Mechanistic studies of the coenzyme F420 reducing formate dehydrogenase from Methanobacterium formicicum. Biochemistry. 1986 Nov 4;25(22):7163–7168. doi: 10.1021/bi00370a059. [DOI] [PubMed] [Google Scholar]
  27. Schauer N. L., Ferry J. G. Metabolism of formate in Methanobacterium formicicum. J Bacteriol. 1980 Jun;142(3):800–807. doi: 10.1128/jb.142.3.800-807.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shuber A. P., Orr E. C., Recny M. A., Schendel P. F., May H. D., Schauer N. L., Ferry J. G. Cloning, expression, and nucleotide sequence of the formate dehydrogenase genes from Methanobacterium formicicum. J Biol Chem. 1986 Oct 5;261(28):12942–12947. [PubMed] [Google Scholar]
  29. Thauer R. K., Jungermann K., Decker K. Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev. 1977 Mar;41(1):100–180. doi: 10.1128/br.41.1.100-180.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tzing S. F., Bryant M. P., Wolfe R. S. Factor 420-dependent pyridine nucleotide-linked formate metabolism of Methanobacterium ruminantium. J Bacteriol. 1975 Jan;121(1):192–196. doi: 10.1128/jb.121.1.192-196.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yamazaki S. A selenium-containing hydrogenase from Methanococcus vannielii. Identification of the selenium moiety as a selenocysteine residue. J Biol Chem. 1982 Jul 25;257(14):7926–7929. [PubMed] [Google Scholar]

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

RESOURCES