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Biodegradation

, Volume 2, Issue 2, pp 71–79 | Cite as

Ether-cleaving enzyme and diol dehydratase involved in anaerobic polyethylene glycol degradation by a new Acetobacterium sp.

  • Edgar Schramm
  • Bernhard Schink
Article

Abstract

A strictly anaerobic, homoacetogenic bacterium was enriched and isolated from anoxic sewage sludge with polyethylene glycol (PEG) 1000 as sole source of carbon and energy, and was assigned to the genus Acetobacterium on the basis of morphological and physiological properties. The new isolate fermented ethylene glycol and PEG's with molecular masses of 106 to 1000 to acetate and small amounts of ethanol. The PEG-degrading activity was not destroyed by proteinase K treatment of whole cells. In cell-free extracts, a diol dehydratase and a PEG-degrading (ether-cleaving) enzyme activity were detected which both formed acetaldehyde as reaction product. The diol dehydratase enzyme was oxygen-sensitive and was stimulated 10–14 fold by added adenosylcobalamine. This enzyme was found mainly in the cytoplasmic fraction (65%) and to some extent (35%) in the membrane fraction. The ether-cleaving enzyme activity reacted with PEG's of molecular masses of 106 to more than 20000. The enzyme was measurable optimally in buffers of high ionic strength (4.0), was extremely oxygen-sensitive, and was inhibited by various corrinoids (adenosylcobalamine, cyanocobalamine, hydroxocobalamine, methylcobalamine). This enzyme was found exclusively in the cytoplasmic fraction. It is concluded that PEG is degraded by this bacterium inside the cytoplasm by a hydroxyl shift reaction, analogous to a diol dehydratase reaction, to form an unstable hemiacetal intermediate. The name polyethylene glycol acetaldehyde lyase is suggested for the responsible enzyme.

Key words

anaerobic degradation ether cleavage reactions corrinoids Acetobacterium sp. polyethylene glycol acetaldehyde lyase 

Abbreviations

EG

ethylene glycol

DiEG

diethylene glycol

TriEG

triethylene glycol

TeEG

tetraethylene glycol

PEG

polyethylene glycol (molecular mass indicated)

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References

  1. Babior BM (1982) Ethanolamine ammonia-lyase. In: Dolphin D (Ed) B12, Vol 2 (pp 264–286). John Wiley and Sons, New YorkGoogle Scholar
  2. Bock KJ & Stache H (1982) Surfactants. In: Hutzinger O (Ed) The Handbook of Environmental Chemistry, Vol 3B (pp 163–199). Springer, BerlinGoogle Scholar
  3. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254Google Scholar
  4. Cox DP (1978) The biodegradation of polyethylene glycols. Adv. Appl. Microbiol. 23: 173–194Google Scholar
  5. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann. NY Acad. Sci. 121: 404–427Google Scholar
  6. Diekert GB & Thauer RK (1978) Carbon monoxide oxidation by Clostridium thermoaceticum and Clostridium formicoaceticum. J. Bacteriol. 136: 597–606Google Scholar
  7. Duine JA, Frank JJ & Jongejan JA (1986) PQQ and quinoprotein enzymes in microbial oxidations. FEMS Microbiol. Rev. 32: 165–178Google Scholar
  8. Dwyer D & Tiedje JM (1983) Degradation of ethylene glycol and polyethylene glycols by methanogenic consortia. Appl Environ. Microbiol. 46: 185–190Google Scholar
  9. Dwyer D & Tiedje JM (1986) Metabolism of polyethylene glycol by two anaerobic bacteria, Desulfovibrio desulfuricans and a Bacteroides sp. Appl. Environ. Microbiol. 52: 852–856Google Scholar
  10. Grant MA & Payne WJ (1983) Anaerobic growth of Alcaligenes faecalis var. denitrificans at the expense of ether glycols and nonionic detergents. Biotechnol. Bioeng. 25: 627–630Google Scholar
  11. Houston CA (1981) Detergent alcohols in the news. J. Am. Oil. Chem. Soc. 58: 873a-874aGoogle Scholar
  12. Haines JR & Alexander M (1975) Microbial degradation of polyethylene glycols. Appl. Microbiol. 29: 621–625Google Scholar
  13. International Union of Biochemistry Nomenclature Committee (Ed) (1984) Enzyme Nomenclature. Academic Press, Orlando FL USAGoogle Scholar
  14. Kawai F (1985) Existence of ether bond-cleaving enzyme in a polyethylene glycol-utilizing symbiotic mixed culture. FEMS Microbiol. Lett. 30: 273–276Google Scholar
  15. Kawai F, Yamanaka H, Ameyama M, Shinagawa E, Matsushita K & Adachi O (1985) Identification of the prosthetic group and further characterization of a novel enzyme, polyethylene glycol dehydrogenase. Agric. Biol. Chem. 49: 1071–1076Google Scholar
  16. Kawai F (1987) The biochemistry of degradation of polyethers. In: CRC Critical Reviews in Biotechnology, Vol 6 (pp 273–307). Chemical Rubber Company Boca Raton FL USAGoogle Scholar
  17. Lugtenberg B, Neijers J, Peters R, van der Hock P & van Alphen L (1975) Electrophoretic resolution of the major outer membrane proteins of Escherichia coli K-12 into four bands. FEBS Lett. 58: 254–255Google Scholar
  18. Magee CM, Rodeheaver G, Edgerton MT & Edlich RF (1975) A more reliable Gram staining technic for diagnosis of surgical infections. Am J Surgery 130: 341–346Google Scholar
  19. Pearce BA & Heydeman MT (1980) Metabolism of di(ethylene glycol) [2-(2′-hydroxyethoxy)ethanol] and other short poly (ethylene glycol)s by Gram-negative bacteria. J. Gen. Microbiol. 118: 21–27Google Scholar
  20. Pfennig N (1978) Rhodocyclus purpureus gen. nov. and ap. nov., a ring-shaped, vitamin B12-requiring member of the family Rhodospirillaceae. Int. J. System. Bacteriol. 23: 283–288Google Scholar
  21. Schink B, Bomar M (1991) The Genera Acetobacterium, Acetogenium, Acetoanaerobium, and Acetitomaculum. In: Balows A, Trüper HG, Dworkin M, Harder W & Schleifer KH (Eds) The Erokaryotes, 2nd edn. Springer Verlag, New York (in press)Google Scholar
  22. Schink B, Pfennig N (1982) Fermentation of trihydroxybenzenes by Pelobacter acidigallici gen. nov. sp. nov., a new strictly anaerobic, non-sporeforming bacterium. Arch. Microbiol. 133: 195–201Google Scholar
  23. Schink B & Stieb M (1983) Fermentative degradation of polyethylene glycol by a strictly anaerobic, Gram-negative, non-sporeforming bacterium, Pelobacter venetianus sp. nov. Appl Environ. Microbiol. 45: 1905–1913Google Scholar
  24. Steber J & Wierich P (1985) Metabolites and biodegradation pathways of fatty alcohol ethoxylates in microbial biocoenoses of sewage treatment plants. Appl. Environ. Microbiol. 49: 530–537Google Scholar
  25. Straß A & Schink B (1986) Fermentation of polyethylene glycol via acetaldehyde in Pelobacter venetianus. Appl. Microbiol. Biotechnol. 25: 37–42Google Scholar
  26. Thélu J, Medina L & Pelmont J (1980) Oxidation of polyoxyethylene oligomers by an inducible enzyme from Pseudomonas P 400. FEMS Microbiol. Lett. 8: 187–190Google Scholar
  27. Toraya T, Honda S & Fukui S (1979) Fermentation of 1,2-Ethanediol by some genera of Enterobacteriaceae involving Coenzyme B12-dependent diol dehydratase. J. Bacteriol. 139: 39–47Google Scholar
  28. Toraya T, Krodel E, Mildvan AS & Abeles H (1979) Role of peripheral side chains of vitamin B12 coenzymes in the reaction catalyzed by dioldehydrase. Biochemistry 18: 417–426Google Scholar
  29. Toraya T & Fukui S (1982) Diol dehydrase. In: Dolphin D (Ed) B12, Vol 2 (pp 233–266). John Wiley and Sons, New YorkGoogle Scholar
  30. Wagener S & Schink B (1988) Fermentative degradation of nonionic surfactants and polyethylene glycol by enrichment cultures and by pure cultures of homoacetogenic and propionate-forming bacteria. Appl. Environ. Microbiol. 54: 561–565Google Scholar
  31. Widdel F & Pfennig N (1981) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments. Description of Desulfobacter postgatei gen. nov. sp. nov. Arch. Microbiol. 134: 286–294Google Scholar
  32. Widdel F, Kohring GW & Mayer F (1983) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. III. Characterization of the filamentous gliding Desulfonema limicola gen. nov. sp. nov., and Desulfonema magnum sp. nov. Arch. Microbiol. 134: 286–294Google Scholar
  33. Zehnder AJB & Wuhrmann K (1976) Titanium-III-citrate as a nontoxic oxidation-reduction-buffering system for the culture of anaerobes. Science 194: 1165–1166Google Scholar

Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Edgar Schramm
    • 1
  • Bernhard Schink
    • 1
  1. 1.Lehrstuhl Mikrobiologie I der Eberhard-Karls-UniversitätTübingenGermany

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