Archives of Microbiology

, Volume 138, Issue 1, pp 72–78 | Cite as

Purification and characterization of an inducible dissimilatory type sulfite reductase from Clostridium pasteurianum

  • Gail Harrison
  • Carol Curle
  • Edward J. Laishley
Original Papers


An inducible sulfite reductase was purified from Clostridium pasteurianum. The pH optimum of the enzyme is 7.5 in phosphate buffer. The molecular weight of the reductase was determined to be 83,600 from sodium dodecyl sulfate gel electrophoresis with a proposed molecular structure: α2β2. Its absorption spectrum showed a maximum at 275 nm, a broad shoulder at 370 nm and a very small absorption maximum at 585 nm. No siroheme chromophore was isolated from this reductase. The enzyme could reduced the following substrates in preferential order: NH2OH> SeO 3 2- >NO 2 2- at rates 50% or less of its preferred substrate SO 3 2- . The proposed dissimilatory intermediates, S3O 6 2- or S2O 3 2- , were not utilized by this reductase while KCN inhibited its activity. Varying the substrate concentration [SO 3 2- ] from 1 to 2.5 μmol affected the stoichiometry of the enzyme reaction by alteration of the ratio of H2 uptake to S2- formed from 2.5:1 to 3.1:1. The inducible sulfite reductase was found to be linked to ferredoxin which could be completely replaced by methyl viologen or partially by benzyl viologen. Some of the above-mentioned enzyme properties and physiological considerations indicated that it was a dissimilatory type sulfite reductase.

Key words

Clostridium posteurianum Sulfite reductase Sulfite metabolism 



sodium dodecyl sulfate


bovine serum albumin


Lactate dehydrogenase


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Akagi JM, Chan M, Adams V (1974) Observations on the bisulfite reductase (P 582) isolated from Desulfotomaculum nigrificans. J Bacteriol 120:240–244Google Scholar
  2. Brewer JM, Pesce AJ, Ashworth RB (1974) Experimental techniques in biochemistry. Prentice-Hall, Inc., Englewood Cliffs, NJ, pp 351–352Google Scholar
  3. Drake HL, Akagi JM (1976) Purification of a unique bisulfite-reducing enzyme from Desulfovibrio vulgaris. Biochem Biophys Res Comm 71:1214–1219Google Scholar
  4. Drake HL, Akagi JM (1977) Bisulfite reductase of Desulfovibrio vulgaris: explanation for product formation. J Bacteriol 32:139–143Google Scholar
  5. Jeng D (1969) Sulfur metabolism of N2-fixing Clostridium pasteurianum. Ph. D. Thesis, Purdue University, West Lafayette, IN, USAGoogle Scholar
  6. Jones HE, Skyring GW (1975) Effect of enzymatic assay conditions on sulfite reduction catalyzed by desulfoviridin from Desulfovibrio gigas. Biochim Biophys Acta 377:52–60Google Scholar
  7. Hatchikian C, Zeikus JG (1983) Characterization of a new type dissimilatory sulfite reductase present in Thermodesulfo-bacterium commune. J Bacteriol 153:1211–1220Google Scholar
  8. Kobayashi K, Takahashi E, Ishimoto M (1972) Biochemical studies on sulfate-reducing bacteria. XI. Purification and some properties of sulfite reductase, desulfoviridin. J Biochem 72:879–887Google Scholar
  9. Kobayashi K, Seki Y, Ishimoto M (1974) Biochemical studies on sulfate-reducing bacteria. XIII. Sulfite reductase from Desulfovibrio vulgaris — mechanism of trithionate, thiosulfate and sulfide formation and enzymatic properties. J Biochem 75:519–529Google Scholar
  10. Laishley EJ, Krouse HR (1978) Stable isotope fractionation by Clostridium pasteurianum. 2. Regulation of sulfite reductases by sulfur amino acids and their influence on sulfur isotope fractionation during SO32- and SO42- reduction. Can J Microbiol 24:716–724Google Scholar
  11. Laishley EJ, Lin P, Peck Jr HD (1971) A ferredoxin-linked sulfite reductase from Clostridium pasteurianum. Can J Microbiol 17:889–895Google Scholar
  12. Laishley EJ, McCready RGL, Bryant R, Krouse HR (1976) Stable isotope fractionation by C. pasteurianum. In: Nriagu JO (ed) Environmental biochemistry, vol I: Carbon, nitrogen, phosphorus, sulfur and selenium cycles. Ann Arbor Science, Ann Arbor, MI, pp 327–349Google Scholar
  13. Laishley E, Tyler M, Krouse H (1984) Sulfur isotope fractionation during SO23 reduction by different clostridial species. Can J Microbiol (in press)Google Scholar
  14. Lee PJ, LeGall J, Peck Jr HD (1973) Isolation of assimilatory and dissimilatory-type sulfite reductases from Desulfovibrio vulgaris. J Bacteriol 115:529–542Google Scholar
  15. Mortenson LE (1964) Purification and analysis of ferredoxin from Clostridium pasteurianum. Biochim Biophys Acta 81:71–77Google Scholar
  16. Murphy MJ, Siegel LM (1973) Siroheme and sirohydrochlorin. The basis for a new type of porphyrin-related prosthetic group common to both assimilatory and dissimilatory sulfite reductases. J Biol Chem 248:6911–6919Google Scholar
  17. Palmer WG (1954) Experimental Inorganic Chemistry. Cambridge University Press, New York, pp 370–371Google Scholar
  18. Peck Jr HD (1967) Some evolutionary aspects of inorganic sulfur metabolism. In: Lectures on theoretical and applied aspects of modern microbiology. University of Maryland Press, College Park, MDGoogle Scholar
  19. Peck Jr HD, Gest H (1956) A new procedure for assay of bacterial hydrogenase. J Bacteriol 71:70–80Google Scholar
  20. Schedel M, Trüper HG (1979) Purification of Thiobacillus denitrificans siroheme sulfite reductase and investigation of some molecular and catalytic properties. Biochim Biophys Acta 568:454–467Google Scholar
  21. Schedel M, Vanselow M, Trüper HG (1979) Siroheme sulfite reductase isolated from Chromatium vinosum: Purification and investigation of some of its molecular and catalytic properties. Arch Microbiol 121:29–36Google Scholar
  22. Schwartz RM, Dayhoff MO (1978) Origins of prokaryotes, cukaryotes, mitochondria and chloroplasts. Science 199: 395–403Google Scholar
  23. Siegel LM, Murphy MJ, Kamin H (1973) Reduced nicotinamide adenine dinucleotide phosphate-sulfite reductase of Enterobacteria. 1. The Escherichia coli hemoflavoprotein: molecular parameters and prosthetic groups. J Biol Chem 248:251–264Google Scholar
  24. Weber K, Osborn M (1969) The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 244:4406–4412Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Gail Harrison
    • 1
  • Carol Curle
    • 1
  • Edward J. Laishley
    • 1
  1. 1.Department of BiologyThe University of CalgaryCalgaryCanada

Personalised recommendations