Characterization of a CO:heterodisulfide oxidoreductase system from acetate-grown Methanosarcina thermophila

C. W. Peer, M. H. Painter, M. E. Rasche, J. G. Ferry

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

During the methanogenic fermentation of acetate by Methanosarcina thermophila, the CO dehydrogenase complex cleaves acetyl coenzyme A and oxidizes the carbonyl group (or CO) to CO2, followed by electron transfer to coenzyme M (CoM)-S-S-coenzyme B (CoB) and reduction of this heterodisulfide to HS-CoM and HS-CoB (A. P. Clements, R. H. White, and J. G. Ferry, Arch. Microbiol. 159:296-300, 1993). The majority of heterodisulfide reductase activity was present in the soluble protein fraction after French pressure cell lysis. A CO:CoM-S-S-CoB oxidoreductase system from acetate-grown cells was reconstituted with purified CO dehydrogenase enzyme complex, ferredoxin, membranes, and partially purified heterodisulfide reductase. Coenzyme F420 (F420) was not required, and CO:F420 oxidoreductase activity was not detected in cell extracts. The membranes contained cytochrome b that was reduced with CO and oxidized with CoM-S-S-CoB. The results suggest that a novel CoM-S-S-CoB reducing system operates during acetate conversion to CH4 and CO2. In this system, ferredoxin transfers electrons from the CO dehydrogenase complex to membrane-bound electron carriers, including cytochrome b, that are required for electron transfer to the heterodisulfide reductase. The cytochrome b was purified from solubilized membrane proteins in a complex with six other polypeptides. The cytochrome was not reduced when the complex was incubated with H2 or CO, and H2 uptake hydrogenase activity was not detected; however, the addition of CO dehydrogenase enzyme complex and ferredoxin enabled the CO-dependent reduction of cytochrome b.

Original languageEnglish (US)
Pages (from-to)6974-6979
Number of pages6
JournalJournal of bacteriology
Volume176
Issue number22
DOIs
StatePublished - Jan 1 1994

Fingerprint

carbon monoxide dehydrogenase
Methanosarcina
Mesna
Carbon Monoxide
Oxidoreductases
Acetates
Cytochromes b
Ferredoxins
Electrons
Membranes
Acetyl Coenzyme A
Enzymes
Cytochromes
Cell Extracts
Fermentation
Membrane Proteins
Pressure
Peptides

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

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title = "Characterization of a CO:heterodisulfide oxidoreductase system from acetate-grown Methanosarcina thermophila",
abstract = "During the methanogenic fermentation of acetate by Methanosarcina thermophila, the CO dehydrogenase complex cleaves acetyl coenzyme A and oxidizes the carbonyl group (or CO) to CO2, followed by electron transfer to coenzyme M (CoM)-S-S-coenzyme B (CoB) and reduction of this heterodisulfide to HS-CoM and HS-CoB (A. P. Clements, R. H. White, and J. G. Ferry, Arch. Microbiol. 159:296-300, 1993). The majority of heterodisulfide reductase activity was present in the soluble protein fraction after French pressure cell lysis. A CO:CoM-S-S-CoB oxidoreductase system from acetate-grown cells was reconstituted with purified CO dehydrogenase enzyme complex, ferredoxin, membranes, and partially purified heterodisulfide reductase. Coenzyme F420 (F420) was not required, and CO:F420 oxidoreductase activity was not detected in cell extracts. The membranes contained cytochrome b that was reduced with CO and oxidized with CoM-S-S-CoB. The results suggest that a novel CoM-S-S-CoB reducing system operates during acetate conversion to CH4 and CO2. In this system, ferredoxin transfers electrons from the CO dehydrogenase complex to membrane-bound electron carriers, including cytochrome b, that are required for electron transfer to the heterodisulfide reductase. The cytochrome b was purified from solubilized membrane proteins in a complex with six other polypeptides. The cytochrome was not reduced when the complex was incubated with H2 or CO, and H2 uptake hydrogenase activity was not detected; however, the addition of CO dehydrogenase enzyme complex and ferredoxin enabled the CO-dependent reduction of cytochrome b.",
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Characterization of a CO:heterodisulfide oxidoreductase system from acetate-grown Methanosarcina thermophila. / Peer, C. W.; Painter, M. H.; Rasche, M. E.; Ferry, J. G.

In: Journal of bacteriology, Vol. 176, No. 22, 01.01.1994, p. 6974-6979.

Research output: Contribution to journalArticle

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T1 - Characterization of a CO:heterodisulfide oxidoreductase system from acetate-grown Methanosarcina thermophila

AU - Peer, C. W.

AU - Painter, M. H.

AU - Rasche, M. E.

AU - Ferry, J. G.

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