Skip to main content
SpringerLink
Log in

Anaerobic energy metabolism of the sulfur-reducing bacterium “Spirillum” 5175 during dissimilatory nitrate reduction to ammonia

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

In a batch culture experiment the microaerophilic Campylobacter-like bacterium “Spirillum” 5175 derived its energy for growth from the reduction of nitrate to nitrite and nitrite to ammonia. Hereby, formate served as electron donor, acetate as carbon source, and l-cysteine as sulfur source. Nitrite was quantitatively accumulated in the medium during the reduction of nitrate; reduction of nitrite began only after nitrate was exhausted from the medium. The molar growth yield per mol formate consumed, Ym, was 2.4g/mol for the reduction of nitrate to nitrite and 2.0 g/mol for the conversion of nitrite to ammonia. The gain of ATP per mol of oxidized formate was 20% higher for the reduction of nitrate to nitrite, compared to the reduction of nitrite to ammonia. With succinate as carbon source and nitrite as electron acceptor, Ym was 3.2g/mol formate, i.e. 60% higher than with acetate as carbon source. No significant amount of nitrous oxide or dinitrogen was produced during growth with nitrate or nitrite both in the presence or absence of acetylene. No growth on nitrous oxide was found. The hexaheme c nitrite reductase of “Spirillum” 5175 was an inducible enzyme. It was present in cells cultivated with nitrate or nitrite as electron acceptor. It was absent in cells grown with fumarate, but appeared in high concentration in “Spirillum” 5175 grown on elemental sulfur. Furthermore, the dissimilatory enzymes nitrate reductase and hexaheme c nitrite reductase were localized in the periplasmic part of the cytoplasmic membrane.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DNRA:

dissimilatory nitrate reduction to ammonia

〈N⦔:

assimilated nitrogen

NaPi :

sodium phosphate

OD578 :

optical density at 578 nm

〈S0〉:

elemental sulfur

“Spirillum” 5175N,F,S :

“Spirillum” 5175 grown with nitrate, fumarate or elemental sulfur as electron acceptor

References

  • Balderston WL, Sherr B, Payne WJ (1976) Blockage by acetylene of nitrous oxide reduction in Pseudomonas perfectomarinus. Appl Environ Microbiol 31: 504–508

    CAS  PubMed  Google Scholar 

  • Barton LL, LeGall J, Odom JM, Peck HDJr (1983) Energy coupling to nitrite respiration in the sulfate-reducing bacterium Desulfovibrio gigas. J Bacteriol 153: 867–871

    CAS  PubMed  Google Scholar 

  • Blümle S, Zumft WG (1991) Respiratory nitrate reductase from denitrifying Pseudomonas stutzeri, purification, properties and target of proteolysis. Biochim Biophys Acta 1057: 102–108

    Google Scholar 

  • Bokranz MJ, Katz J, Schröder I, Roberton AM, Kröger A (1983) Energy metabolism and biosynthesis of Vibrio succinogenes growing with nitrate or nitrite as terminal electron acceptor. Arch Microbiol 135: 36–41

    Article  CAS  Google Scholar 

  • Boltz DF, Taras MJ (1978) Nitrogen. In: Boltz DF, Howell JA (eds) Colorimetric determination of nonmetals. Chemical analysis, vol 8, 2nd edn. John Wiley & Sons, New York Chichester Brisbane, Toronto, pp 197–251

    Google Scholar 

  • Bronder M, Mell H, Stupperich E, Kröger A (1982) Biosynthetic pathways of Vibrio succinogenes growing with fumarate as terminal electron acceptor and sole carbon source. Arch Microbiol 131: 216–223

    Article  CAS  PubMed  Google Scholar 

  • Cline JD (1969) Spectrophotometric determination of hydrogen sulfide in natural waters. Limnol Oceanogr 14: 454–458

    CAS  Google Scholar 

  • Cole JA (1988) Assimilatory and dissimilatory reduction of nitrate to ammonia. In: Cole JA, Ferguson SJ (eds) The nitrogen and sulphur cycles. Society for General Microbiology, Cambridge University Press, Cambridge. pp 281–329

    Google Scholar 

  • Conrad R, Seiler W (1980) Field measurements of the loss of fertilizer nitrogen into the atmosphere as nitrous oxide. Atmos Environ 14: 555–558

    CAS  Google Scholar 

  • Coyne MS, Arunakumari A, Pankratz HS, Tiedje JM (1990) Localization of the cytochrome cd 1 and copper nitrite reductases in denitrifying bacteria. J Bacteriol 172: 2558–2562

    CAS  PubMed  Google Scholar 

  • Cypionka H, Pfennig N (1986) Growth yields of Desulfotomaculum orientis with hydrogen in chemostat culture. Arch Microbiol 143: 396–399

    Article  CAS  Google Scholar 

  • Denis KS, Dias FM, Rowe JJ (1990) Oxygen regulation of nitrate transport by diversion of electron flow in Escherichia coli. J Biol Chem 265: 18095–18097

    CAS  PubMed  Google Scholar 

  • Fujita T, Sato R (1966) Studies on soluble cytochromes in Enterobacteriaceae III. Localization of cytochrome c 552 in the surface layer of cells. J Biochem 60: 568–577

    CAS  Google Scholar 

  • Heppel LA (1969) The effect of osmotic shock on release of bacterial proteins and on active transport. J Gen Physiol 54: 95–109

    Article  Google Scholar 

  • Jackson RH, Cornish-Bowden A, Cole JA (1981) Prosthetic groups of the NADH-dependent nitrite reductase from Escherichia coli K12. Biochem J 193: 861–867

    CAS  PubMed  Google Scholar 

  • Jones RW, Garland PB (1977) Sites and specificity of the reaction of bipyridylium compounds with anaerobic respiratory enzymes of Escherichia coli. Effects of permeability barriers imposed by the cytoplasmic membrane. Biochem J 164: 199–211

    CAS  PubMed  Google Scholar 

  • Koike I, Hattori A (1975) Energy yield of denitrification: an estimate from growth yield in continuous cultures of Pseudomonas denitrificans under nitrate-, nitrite- and nitrous oxidelimited conditions. J Gen Microbiol 88: 11–19

    CAS  PubMed  Google Scholar 

  • Kröger A, Dorrer E, Winkler E (1980) The orientation of the substrate sites of formate dehydrogenase and fumarate reductase in the membrane of Vibrio succinogenes. Biochim Biophys Acta 589: 118–136

    PubMed  Google Scholar 

  • Laanbroek HJ, Kingma W, Veldkamp H (1977) Isolation of an aspartate-fermenting, free-living Campylobacter species. FEMS Microbiol Lett 1: 99–102

    Article  CAS  Google Scholar 

  • Lang E, Lang H (1972) Spezifische Farbreaktion zum direkten Nachweis der Ameisensäure. Z Anal Chem 260: 8–10

    Article  CAS  Google Scholar 

  • Liu M-C, Peck HDJr (1981) The isolation of a hexaheme cytochrome from Desulfovibrio desulfuricans and its identification as a new type of nitrite reductase. J Biol Chem 256: 13159–13164

    CAS  PubMed  Google Scholar 

  • Luria SE (1960) The bacterial protoplasm: composition and organization. In: Gunsalus IC, Stanier RY (eds) The bacteria, vol I. Academic Press, New York, pp 1–34

    Google Scholar 

  • McEwan AG, Jackson JB, Ferguson SJ (1984) Rationalization of properties of nitrate reductases in Rhodopseudomonas capsulata. Arch Microbiol 137: 344–349

    Article  CAS  Google Scholar 

  • Pope NR, Cole JA (1982) Generation of a membrane potential by one of two independent pathways for nitrite reduction by Escherichia coli. J Gen Microbiol 128: 319–322

    Google Scholar 

  • Schumacher W, Kroneck PMH (1991) Dissimilatory hexaheme c nitrite reductase of “Spirillum” strain 5175: purification and properties. Arch Microbiol 156: 70–74

    Article  CAS  Google Scholar 

  • Seitz H-J, Cypionka H (1986) Chemolithotrophic growth of Desulfovibrio desulfuricans with hydrogen coupled to ammonification of nitrate or nitrite. Arch Microbiol 146: 63–67

    Article  CAS  Google Scholar 

  • Skirrow G (1965) The dissolved gases-carbon dioxide. In: Riley JP, Skirrow G (eds) Chemical oceanography, vol I. Academic Press, London, p. 227

    Google Scholar 

  • Smith AF (1974) Malate to oxaloacetate reaction. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol III. Verlag Chemie, Weinheim Deerfield Beach Basel, pp 166–171

    Google Scholar 

  • Smith MS (1983) Nitrous oxide production by Escherichia coli is correlated with nitrate reductase activity. Appl Environ Microbiol 45: 1545–1547

    CAS  PubMed  Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150: 76–85

    Article  CAS  PubMed  Google Scholar 

  • Steenkamp DJ, Peck HD (1981) Proton translocation associated with nitrite respiration in Desulfovibrio desulfuricans. J Biol Chem 256: 5450–5458

    CAS  PubMed  Google Scholar 

  • Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41: 100–180

    CAS  PubMed  Google Scholar 

  • Tiedje JM (1988) Ecology of denitrification and dissimilatory nitrate reduction to ammonium. In: Zehnder AJB (ed) Biology of anaerobic microorganisms. John Wiley & Sons, New York Chichester Brisbane, pp 179–244

    Google Scholar 

  • Vries Wde, Niekus HGD, Boellaard M, Stouthamer AH (1980) Growth yields and energy generation by Campylobacter sputorum subspecies bubulus during growth in continuous culture with different hydrogen acceptors. Arch Microbiol 124: 221–227

    Article  PubMed  Google Scholar 

  • Vries Wde, Niekus HGD, Berchum Hvan, Stouthamer AH (1982) Electron transport-linked proton translocation at nitrite reduction in Campylobacter sputorum subspecies bubulus. Arch Microbiol 131: 132–139

    Article  PubMed  Google Scholar 

  • Wolfe RS, Pfennig N (1977) Reduction of sulfur by spirillum 5175 and syntrophism with Chlorobium. Appl Environ Microbiol 33: 427–433

    CAS  PubMed  Google Scholar 

  • Yoshinari T (1980) N2O Reduction by Vibrio succinogenes. Appl Environ Microbiol 39: 81–84

    CAS  PubMed  Google Scholar 

  • Zöphel A, Kennedy MC, Beinert H, Kroneck PMH (1988) Investigations on microbial sulfur respiration. 1. Activation and reduction of elemental sulfur in several strains of eubacteria. Arch Microbiol 150: 72–77

    Article  Google Scholar 

  • Zöphel A, Kennedy MC, Beinert H, Kroneck PMH (1991) Investigations on microbial sulfur respiration. 2. Isolation, purification, and characterization of cellular components from Spirillum 5175. Eur J Biochem 195: 849–856

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Fakultät für Biologie, Universität Konstanz, Universitätsstrasse 10, W-7750, Konstanz, Federal Republic of Germany

    Wolfram Schumacher & Peter M. H. Kroneck

Authors
  1. Wolfram Schumacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter M. H. Kroneck
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schumacher, W., Kroneck, P.M.H. Anaerobic energy metabolism of the sulfur-reducing bacterium “Spirillum” 5175 during dissimilatory nitrate reduction to ammonia. Arch. Microbiol. 157, 464–470 (1992). https://doi.org/10.1007/BF00249106

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00249106

Key words

Search

Navigation