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Veratrol-O-demethylase of Acetobacterium dehalogenans: ATP-dependent reduction of the corrinoid protein

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Abstract

The anaerobic veratrol O-demethylase mediates the transfer of the methyl group of the phenyl methyl ether veratrol to tetrahydrofolate. The primary methyl group acceptor is the cobalt of a corrinoid protein, which has to be in the +1 oxidation state to bind the methyl group. Due to the negative redox potential of the cob(II)/cob(I)alamin couple, autoxidation of the cobalt may accidentally occur. In this study, the reduction of the corrinoid to the superreduced [CoI] state was investigated. The ATP-dependent reduction of the corrinoid protein of the veratrol O-demethylase was shown to be dependent on titanium(III) citrate as electron donor and on an activating enzyme. In the presence of ATP, activating enzyme, and Ti(III), the redox potential versus the standard hydrogen electrode (E SHE) of the cob(II)alamin/cob(I)alamin couple in the corrinoid protein was determined to be −290 mV (pH 7.5), whereas E SHE at pH 7.5 was lower than −450 mV in the absence of either activating enzyme or ATP. ADP, AMP, or GTP could not replace ATP in the activation reaction. The ATP analogue adenosine-5′-(β,γ-imido)triphosphate (AMP-PNP, 2–4 mM) completely inhibited the corrinoid reduction in the presence of ATP (2 mM).

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Abbreviations

Odm :

O-demethylase

Odm ver :

Veratrol O-demethylase

CPver :

Corrinoid protein of the veratrol O-demethylase

AEver :

Activating enzyme of the veratrol O-demethylase

AE:

Activating enzyme of the O-demethylases

MAP:

Methanol:coenzyme M methyltransferase activating protein

MTIver :

Methyltransferase I of the veratrol O-demethylase (veratrol:corrinoid protein methyltransferase)

H4F:

Tetrahydrofolate

AMP-PNP:

Adenosin-5′-(β,γ-imido)triphosphate)

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Acknowledgements

This work was supported by grants from the Deutsche Forschungsgemeinschaft and the European Commission. We thank Dr. Dirk Merten (Institute of Applied Geology, Friedrich-Schiller-University Jena) for performing the cobalt analysis.

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  1. Institut für Mikrobiologie, FSU Jena, Lehrstuhl für Angewandte und Ökologische Mikrobiologie, Philosophenweg 12, 07743, Jena, Germany

    Anke Siebert, Torsten Schubert, Tina Engelmann, Sandra Studenik & Gabriele Diekert

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  1. Anke Siebert
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  2. Torsten Schubert
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  3. Tina Engelmann
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Correspondence to Gabriele Diekert.

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Siebert, A., Schubert, T., Engelmann, T. et al. Veratrol-O-demethylase of Acetobacterium dehalogenans: ATP-dependent reduction of the corrinoid protein. Arch Microbiol 183, 378–384 (2005). https://doi.org/10.1007/s00203-005-0001-8

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