Abstract
The pathway of anaerobic acetone degradation by the denitrifying bacterial strain BunN was studied by enzyme measurements in extracts of anaerobic acetone-grown cells. An ADP- and MgCl2-dependent decarboxylation of acetoacetate was detected which could not be found in cell-free extracts of acetate-grown cells. It is concluded that free acetoacetate is formed by ATP-dependent carboxylation of acetone. Acetoacetate was converted into its coenzyme A ester by succinyl-CoA: acetoacetate CoA transferase, and cleaved by a thiolase into acetyl-CoA. The acetyl residue was completely oxidized in the citric acid cycle. The ADP-dependent decarboxylation of acetoacetate was inhibited by EDTA, but not by avidin. High myokinase activities led to equilibrium amounts of ATP, ADP, and AMP in the reaction mixtures, and prevented determination of the decarboxylase reaction stoichiometry, therefore.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ADP:
-
adenosine diphosphate
- AMP:
-
adenosine monophosphate
- ATP:
-
adenosine triphosphate
- BSA:
-
bovine serum albumine
- MOPS:
-
3-(N-morpholino)propanesulfonic acid
- PIPES:
-
piperazine-N,N′-bis-(2-ethanesulfonic acid)
- PHB:
-
poly-β-hydroxybutyrate
- Tris:
-
Tris-(hydroxymethyl-) aminomethane
References
Bergmeyer HU (Ed) (1983) Methods of Enzymatic Analysis, Vol I–III. Verlag Chemie, Weinheim
Boehringer GmbH (1973) Biochemica Information I (p 53). Mannheim
Bonnet-Smits EM, Robertson LA, Van Dijken JP, Senior E & Kuenen JG (1988) Carbon dioxide fixation as the initial step in the metabolism of acetone by Thiosphaera pantotropha. J. Gen. Microbiol. 134: 2281–2289
Cooper RA & Kornberg HL (1974) Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase. In: Boyer PD (Ed), The Enzymes, Vol X (pp 631–649). Academic Press, New York
Dixon GH & Kornberg HL (1959) Assay methods for key enzymes of the glyoxylate cycle. Biochem. J. 72: 3P
Fridovich I (1972) Acetoacetate decarboxylase. In: Boyer PD (Ed) The Enzymes, Vol VI, third edition: Carboxylation and decarboxylation (nonoxidative), Isomerisation (pp 117–168). Academic Press, New York
Hall LM (1962) Preparation of crystalline lithium acetoacetate. Analyt. Biochem. 3: 75–80
Herbert D, Phipps PJ & Strange RE (1971) Chemical analysis of microbial cells. In: Norris JR & Ribbons DW (Eds) Methods in Microbiology, Vol 5B (pp 209–344). Academic Press, New York
International union of biochemistry. Nomenclature committee (Ed) (1984) Enzyme nomenclature. Academic Press, Orlando
Lehninger AL (1983) Biochemie, second edition (p 679). Verlag Chemie, Weinheim
Lukins HB & Foster JW (1963) Methylketone metabolism in hydrocarbon utilizing mycobacteria. J. Bacteriol. 85: 1074–1087
Platen H (1989) Abbau von Aceton und höheren aliphatischen Ketonen durch anaerobe Bakterien. Thesis, Universität Tübingen, FRG
Platen H & Schink B (1987) Methanogenic degradation of acetone by an enrichment culture. Arch. Microbiol. 149: 136–141
Platen H & Schink B (1989) Anaerobic degradation of acetone and higher ketones via carboxylation by newly isolated denitrifying bacteria. J. Gen. Microbiol. 135: 883–891
Rast D & Bachofen R (1967) Carboxylierungsreaktionen in Agaricus bisporus. II. Aceton als ein CO2-Acceptor. Arch. Mikrobiol. 58: 339–167
Rossi A (1938) Azione in vitro del sangue sull' acido acetacetico e β-ossibutirrico. Arch. Sci. Biol. (Bologna) 24: 73–82
Rudney H (1954) Propanediol phosphate as a possible intermediate in the metabolism of acetone. J. Biol. Chem. 210: 361–371
Scrutton MC & Young MR (1972) Pyruvate carboxylase. In: Boyer PD (Ed) The Enzymes, Vol VI, third edition: Carboxylation and Decarboxylation (nonoxidative), Isomerisation (pp 1–35). Academic Press, New York
Senior PJ & Dawes EA (1973) The regulation of poly-β-hydroxybutyrate metabolism in Azotobacter beijerinckii. Biochem. J. 134: 225–238
Siegel JM (1950) The metabolism of acetone by the phostosynthetic bacterium Rhodopseudomonas gelatinosa. J. Bacteriol. 60: 595–606
Stern JR (1956) Optical properties of acetoacetyl-S-coenzyme A and its metal chelates. J. Biol. Chem. 221: 33–44
Taylor DG, Trudgill PW, Gripps RE & Harris PR (1980) The microbial metabolism of acetone. J. Gen. Microbiol. 118: 159–170
Thauer RK, Jungermann K & Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol. Rev. 41: 100–180
Utter MF & Kolenbrander HM (1972) Formation of oxaloacetate by CO2 fixation of phosphoenolpyruvate. In: Boyer PD (Ed) The Enzymes, Vol VI, third edition: Carboxylation and Decarboxylation (nonoxidative), Isomerisation (pp 117–168). Academic Press, New York
Williamson DH, Mellanby J & Krebs HA (1962) Enzymatic determination of D(-)-β-hydroxybutyric acid and acetoacetic acid in blood. Biochem. J. 82: 90–96
Zamenhoff S (1957) Preparation and assay of deoxyribonucleic acid from animal tissue. In: Colowick SP, Kaplan NO (Eds): Methods in Enzymology, Vol III (pp 696–704). Academic Press, New York
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Platen, H., Schink, B. Enzymes involved in anaerobic degradation of acetone by a denitrifying bacterium. Biodegradation 1, 243–251 (1990). https://doi.org/10.1007/BF00119761
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00119761