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Anaerobic acetate oxidation to CO2 by Desulfotomaculum acetoxidans

Demonstration of enzymes required for the operation of an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway

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Abstract

Desulfotomaculum acetoxidans has been proposed to oxidize acetate to CO2 via an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway rather than via the citric acid cycle. We report here the presence of the enzyme activities required for the operation of the novel pathway. In cell extracts the following activities were found (values in brackets=specific activities and apparent K m; 1 U·mg-1=1 μmol·min-1·mg protein-1 at 37°C): Acetate kinase (6.3 U·mg-1; 2 mM acetate; 2.4 mM ATP); phosphate acetyltransferase (60 U·mg-1, 0.4 mM acetylphosphate; 0.1 mM CoA); carbon monoxide dehydrogenase (29 U·mg-1; 13% carbon monoxide; 1.3 mM methyl viologen); 5,10-methylenetetrahydrofolate reductase (3 U·mg-1, 0.06 mM CH3−FH4); methylenetetrahydrofolate dehydrogenase (3.6 U·mg-1, 0.9 mM NAD, 0.1 mM CH2=FH4); methenyltetrahydrofolate cyclohydrolase (0.3 U·mg-1); formyltetrahydrofolate synthetase (3 U·mg-1, 1.4 mM FH4, 0.4 mM ATP, 13 mM formate); and formate dehydrogenase (10 U·mg-1, 0.4 mM formate, 0.5 mM NAD). The specific activities are sufficient to account for the in vivo acetate oxidation rate of 0.26 U·mg-1.

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Abbreviations

FH4 :

Tetrahydrofolate

CHO-FH4 :

N10-formyltetrahydrofolate

CH≡FH4 :

N5,N10-methenyltetrahydrofolate

CH2=FH4 :

N5,N10-methylenetetrahydrofolate

CH3−FH4 :

N5-methyltetrahydrofolate

MOPS:

morpholinopropane sulfonic acid

DTT:

d,l-1,4-dithiothreitol

TRIS:

tris-(hydroxymethyl)-aminomethane

Ap5A:

p1,P5-di(adenosine-5′)pentaphosphate

MV:

methyl viologen

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  1. Mikrobiologie, Fachbereich Biologie, Philipps-Universität, Karl-von-Frisch-Straße, D-3550, Marburg, Federal Republic of Germany

    Alfred M. Spormann & Rudolf K. Thauer

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  1. Alfred M. Spormann
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  2. Rudolf K. Thauer
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Spormann, A.M., Thauer, R.K. Anaerobic acetate oxidation to CO2 by Desulfotomaculum acetoxidans . Arch. Microbiol. 150, 374–380 (1988). https://doi.org/10.1007/BF00408310

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  • DOI: https://doi.org/10.1007/BF00408310

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