Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jan;173(2):929–932. doi: 10.1128/jb.173.2.929-932.1991

Demonstration of carbon-carbon bond cleavage of acetyl coenzyme A by using isotopic exchange catalyzed by the CO dehydrogenase complex from acetate-grown Methanosarcina thermophila.

S A Raybuck 1, S E Ramer 1, D R Abbanat 1, J W Peters 1, W H Orme-Johnson 1, J G Ferry 1, C T Walsh 1
PMCID: PMC207094  PMID: 1987173

Abstract

The purified nickel-containing CO dehydrogenase complex isolated from methanogenic Methanosarcina thermophila grown on acetate is able to catalyze the exchange of [1-14C] acetyl-coenzyme A (CoA) (carbonyl group) with 12CO as well as the exchange of [3'-32P]CoA with acetyl-CoA. Kinetic parameters for the carbonyl exchange have been determined: Km (acetyl-CoA) = 200 microM, Vmax = 15 min-1. CoA is a potent inhibitor of this exchange (Ki = 25 microM) and is formed under the assay conditions because of a slow but detectable acetyl-CoA hydrolase activity of the enzyme. Kinetic parameters for both exchanges are compared with those previously determined for the acetyl-CoA synthase/CO dehydrogenase from the acetogenic Clostridium thermoaceticum. Collectively, these results provide evidence for the postulated role of CO dehydrogenase as the key enzyme for acetyl-CoA degradation in acetotrophic bacteria.

Full text

PDF
931

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abbanat D. R., Ferry J. G. Synthesis of acetyl coenzyme A by carbon monoxide dehydrogenase complex from acetate-grown Methanosarcina thermophila. J Bacteriol. 1990 Dec;172(12):7145–7150. doi: 10.1128/jb.172.12.7145-7150.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Diekert G., Ritter M. Purification of the nickel protein carbon monoxide dehydrogenase of Clostridium thermoaceticum. FEBS Lett. 1983 Jan 10;151(1):41–44. doi: 10.1016/0014-5793(83)80338-x. [DOI] [PubMed] [Google Scholar]
  3. Lu W. P., Harder S. R., Ragsdale S. W. Controlled potential enzymology of methyl transfer reactions involved in acetyl-CoA synthesis by CO dehydrogenase and the corrinoid/iron-sulfur protein from Clostridium thermoaceticum. J Biol Chem. 1990 Feb 25;265(6):3124–3133. [PubMed] [Google Scholar]
  4. Pezacka E., Wood H. G. The autotrophic pathway of acetogenic bacteria. Role of CO dehydrogenase disulfide reductase. J Biol Chem. 1986 Feb 5;261(4):1609–1615. [PubMed] [Google Scholar]
  5. Ragsdale S. W., Wood H. G. Acetate biosynthesis by acetogenic bacteria. Evidence that carbon monoxide dehydrogenase is the condensing enzyme that catalyzes the final steps of the synthesis. J Biol Chem. 1985 Apr 10;260(7):3970–3977. [PubMed] [Google Scholar]
  6. Ragsdale S. W., Wood H. G., Antholine W. E. Evidence that an iron-nickel-carbon complex is formed by reaction of CO with the CO dehydrogenase from Clostridium thermoaceticum. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6811–6814. doi: 10.1073/pnas.82.20.6811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ramer S. E., Raybuck S. A., Orme-Johnson W. H., Walsh C. T. Kinetic characterization of the [3'-32P]coenzyme A/acetyl coenzyme A exchange catalyzed by a three-subunit form of the carbon monoxide dehydrogenase/acetyl-CoA synthase from Clostridium thermoaceticum. Biochemistry. 1989 May 30;28(11):4675–4680. doi: 10.1021/bi00437a025. [DOI] [PubMed] [Google Scholar]
  8. Raybuck S. A., Bastian N. R., Orme-Johnson W. H., Walsh C. T. Kinetic characterization of the carbon monoxide-acetyl-CoA (carbonyl group) exchange activity of the acetyl-CoA synthesizing CO dehydrogenase from Clostridium thermoaceticum. Biochemistry. 1988 Oct 4;27(20):7698–7702. doi: 10.1021/bi00420a019. [DOI] [PubMed] [Google Scholar]
  9. Terlesky K. C., Barber M. J., Aceti D. J., Ferry J. G. EPR properties of the Ni-Fe-C center in an enzyme complex with carbon monoxide dehydrogenase activity from acetate-grown Methanosarcina thermophila. Evidence that acetyl-CoA is a physiological substrate. J Biol Chem. 1987 Nov 15;262(32):15392–15395. [PubMed] [Google Scholar]
  10. Terlesky K. C., Nelson M. J., Ferry J. G. Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila. J Bacteriol. 1986 Dec;168(3):1053–1058. doi: 10.1128/jb.168.3.1053-1058.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Thauer R. K., Möller-Zinkhan D., Spormann A. M. Biochemistry of acetate catabolism in anaerobic chemotrophic bacteria. Annu Rev Microbiol. 1989;43:43–67. doi: 10.1146/annurev.mi.43.100189.000355. [DOI] [PubMed] [Google Scholar]
  12. Wood H. G., Ragsdale S. W., Pezacka E. A new pathway of autotrophic growth utilizing carbon monoxide or carbon dioxide and hydrogen. Biochem Int. 1986 Mar;12(3):421–440. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES