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Archives of Microbiology

, Volume 159, Issue 4, pp 345–353 | Cite as

Anaerobic transformation of 2,4,6-trinitrotoluene (TNT)

  • Andrea Preuss
  • Jürgen Fimpel
  • Gabriele Diekert
Original Papers

Abstract

A sulfate-reducing bacterium using trinitrotoluene (TNT) as the sole nitrogen source was isolated with pyruvate and sulfate as the energy sources. The organism was able to reduce TNT to triaminotoluene (TAT) in growing cultures and cell suspensions and to further transform TAT to still unknown products. Pyruvate, H2, or carbon monoxide served as the electron donors for the reduction of TNT. The limiting step in TNT conversion to TAT was the reduction of 2,4-diamino-6-nitrotoluene (2,4-DANT) to triaminotoluene. The reduction proceeded via 2,4-diamino-6-hydroxylaminotoluene (DAHAT) as an intermediate. The intermediary formation of DAHAT was only observed in the presence of carbon monoxide or hydroxylamine, respectively. The reduction of DAHAT to triaminotoluene was inhibited by both CO and NH2OH. The inhibitors as well as DANT and DAHAT significantly inhibited sulfide formation from sulfite. The data were taken as evidence for the involvement of dissimilatory sulfite reductase in the reduction of DANT and/or DAHAT to triaminotoluene. Hydrogenase purified from Clostridium pasteurianum and carbon monoxide dehydrogenase partially purified from Clostridium thermoaceticum also catalyzed the reduction of DANT in the presence of methyl viologen or ferredoxin, however, as the main reduction product DAHAT rather than triaminotoluene was formed. The findings could explain the function of CO as an electron donor for the DANT reduction (to DAHAT) and the concomitant inhibitory effect of CO on triaminotoluene formation (from DAHAT) by the inhibition of sulfite reductase. Triaminotoluene is further anaerobically converted to unknown products by the isolate under sulfate-reducing and by a Pseudomonas strain under denitrifying conditions. Triaminotoluene conversion was also catalyzed in the absence of cells under aerobic conditions by trace elements, especially by Mn2+, accompanied by the elimination of ammonia in a stoichiometry of 1 NH3 released per TAT transformed. The results might be of interest for the bioremediation of wastewater polluted with nitroaromatic compounds.

Key words

TNT degradation Polynitroaromatic compounds Sulfite reductase Carbon monoxide dehydrogenase Hydrogenase Ferredoxin Sulfidogenic bacteria 

Abbreviations

TNT =

2,4,6-Trinitrotoluene DANT

2,4-DANT =

2,4-Diamino-6-nitrotoluene

2,6-DANT =

2,6-Diamino-4-nitrotoluene

ADNT =

aminodinitrotoluene

2-ADNT and 4-ADNT

amino substituent at positions 2 or 4

TAT =

2,4,6-Triaminotoluene

DAHAT =

2,4-Diamino-6-hydroxylaminotoluene

MV =

Methyl viologen

Fd =

Ferredoxin

H2ase =

Hydrogenase

CODH =

Carbon monoxide dehydrogenase

Pyr: Fd OR =

Pyruvate: ferredoxin oxidoreductase

U =

Units = μmol of substrate converted per min

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References

  1. Amerkhanova NN, Naumova RP (1978) 2,4,6-Trinitrotoluene as a source of nutrition for bacteria. Microbiologiya 47: 393–395Google Scholar
  2. Angermaier L, Simon H (1983) On the reduction of aliphatic and aromatic nitro compounds by clostridia, the role of ferredoxin and its stabilization. Hoppe-Seyler's Z Physiol Chem 365: 961–975Google Scholar
  3. Balch WE, Schoberth S, Tanner RS, Wolfe RS (1977) Acetobacterium, a new genus of hydrogen-oxidizing, carbon-dioxide-reducing anaerobic bacteria. Int J Syst Bacteriol 27: 355–361Google Scholar
  4. Bergmeyer HU, Beutler HO (1985) Ammonia. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Verlag Chemie, Weinheim, 3rd edn vol. VIII, pp 454–461Google Scholar
  5. Boopathy R, Kulpa CF (1992) Trinitrotoluene (TNT) as a sole nitrogen source for a sulfate-reducing bacterium Desulfovibrio sp. (B strain) isolated from an anaerobic digestor. Curr Microbiol 25: 235–241Google Scholar
  6. Braun H, Schmidtchen FP, Schneider A, Simon H (1991) Microbial reduction of N-allylhydroxylamines to N-allylamines using clostridia. Tetrahedron 47: 3329–3334Google Scholar
  7. Cline JD (1969) Spectrophotometric determination of hydrogensulfide in natural waters. Limnol Oceanogr 14: 454–458Google Scholar
  8. Diekert GB, Graf EG, Thauer RK (1979) Nickel requirement for carbon monoxide dehydrogenase formation in Clostridium pasteurianum. Arch Microbiol 122: 117–120Google Scholar
  9. Diekert G, Ritter M (1983) Purification of the nickel protein carbon monoxide dehydrogenase of Clostridium thermoaceticum. FEBS Lett 151: 41–44Google Scholar
  10. Diekert GB, Thauer RK (1978) Carbon monoxide oxidation by Clostridium thermoaceticum and Clostridium formicoaceticum. J Bacteriol 136: 597–606Google Scholar
  11. Dorn M, Andreesen JR, Gottschalk G (1978) Fermentation of fumarate and l-malate by Clostridium formicoaceticum. J Bacteriol 133: 26–32Google Scholar
  12. Fernando T, Bumpus JA, Aust SD (1990) Biodegradation of TNT (2,4,6-trinitrotoluene) by Phanerochaete chrysosporium. Appl Environ Microbiol 56: 1666–1671Google Scholar
  13. Haas R, vonLöw E (1986) Grundwasserbelastung durch eine Altlast. Die Folgen einer ehemaligen Sprengstoffproduktion für die heutige Trinkwassergewinnung. Forum Städtehygiene 37: 33–43Google Scholar
  14. Hallas LE, Alexander M (1983) Microbial transformation of nitroaromatic compounds in sewage effluent. Appl Environ Microbiol 45: 1234–1241Google Scholar
  15. Houben-Weyl (1977) Methoden der Organischen Chemie, vol. XI/1, 4th edn. Thieme, StuttgartGoogle Scholar
  16. Kaplan DL, Kaplan AM (1982) Thermophilic biotransformations of 2,4,6-trinitrotoluene under simulated composting conditions. Appl Environ Microbiol 44: 757–760Google Scholar
  17. Keller C (1992) Biologischer Abbau von TNT. Bioforum 15: 178Google Scholar
  18. LeGall J, Fauque G (1988) Dissimilatory reduction of sulfur compounds. In: Zehnder AJB (ed) Biology of anaerobic microorganisms. Wiley, New York, pp 587–640Google Scholar
  19. Lindemann H (1928) Zum Abbau der Säureazide nach Curtius. Helv Chim Acta 11: 1027–1028Google Scholar
  20. McCormick NG, Feeherry FE, Levinson HS (1976) Microbial transformation of 2,4,6-trinitrotoluene and other nitroaromatic compounds. Appl Environ Microbiol 31: 949–958Google Scholar
  21. Meier-Schneiders M, Busch C, Diekert G (1993) The attachment of bacterial cells to surfaces under anaerobic conditions. Appl Microbiol Biotechnol (in press)Google Scholar
  22. Naumova RP, Selivanovskaya SY, Cherepneva IE (1989) Conversion of 2,4,6-trinitrotoluene under conditions of oxygen and nitrate respiration of Pseudomonas fluorescens. Appl Biochem Microbiol 24: 409–413Google Scholar
  23. Neumeier W, Haas R, Löw Evon (1989) Mikrobieller Abbau von Nitroaromaten aus einer ehemaligen Sprengstoffproduktion. Forum Städte-Hygiene 40: 32–37Google Scholar
  24. O'Brien RW, Morris JG (1971) The Ferredoxin-dependent reduction of chloramphenicol by Clostridium acetobutylicum. J Gen Microbiol 67: 265–271Google Scholar
  25. Parrish FW (1977) Fungal transformation of 2,4-dinitrotoluene and 2,4,6-trinitrotoluene. Appl Environ Microbiol 34: 232–233Google Scholar
  26. Ruggli P, Zaeslin H (1936) Über zwei neue Dichlor-o-nitro-benzoesäuren. Helv Chim Acta 19: 434–439Google Scholar
  27. Schackmann A, Müller R (1991) Reduction of nitroaromatic compounds by different Pseudomonas species under aerobic conditions. Appl Microbiol Biotechnol 34: 809–813Google Scholar
  28. Schnell S, Schink B (1991) Anaerobic anilene degradation via reductive deamination of 4-amino-benzoyl-CoA in Desulfobacterium anilini. Arch Microbiol 155: 183–190Google Scholar
  29. Schönheit P, Wäscher C, Thauer RK (1978) A rapid procedure for the purification of ferredoxin from clostridia using polyethylenimine. FEBS Lett 89: 219–222Google Scholar
  30. Widdel F, Hansen TA (1992) The dissimilatory sulfate- and sulfur-reducing bacteria. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes, vol. I. Springer, Berlin Heidelberg New York, pp 583–624Google Scholar
  31. Wolin EA, Wolin MJ, Wolfe RS (1963) Fermentation of methane by bacterial extracts. J Biol Chem 238: 2882–2886Google Scholar
  32. Zeikus JG, Fuchs G, Kenealy W, Thauer RK (1977) Oxidoreductases involved in cell carbon synthesis of Methanobacterium thermoautotrophicum. J Bacteriol 132: 604–613Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Andrea Preuss
    • 1
  • Jürgen Fimpel
    • 1
  • Gabriele Diekert
    • 1
  1. 1.Institut für MikrobiologieUniversität StuttgartStuttgart 1Germany

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