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Methyltetrahydromethanopterin as an intermediate in methanogenesis from acetate in Methanosarcina barkeri

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Abstract

Cell extracts (100,000×g) of acetate grown Methanosarcina barkeri (strain MS) catalyzed CH4 and CO2 formation from acetyl-CoA with specific activities of 50 nmol·min-1·mg protein-1. CH4 formation was found to be dependent on tetrahydromethanopterin (H4MPT) (apparent K M=4 μM), coenzyme M (H-S-CoM), and 7-mercaptoheptanoylthreonine phosphate (H-S-HTP=component B) rather than on methanofuran (MFR) and coenzyme F420 (F420). Methyl-H4MPT was identified as an intermediate. This compound accumulated when H-S-CoM and H-S-HTP were omitted from the assays. These and previous results indicate that methanogenesis from acetate proceeds via acetyl phosphate, acetyl-CoA, methyl-H4MPT, and CH3-S-CoM as intermediates. The disproportionation of formaldehyde to CO2 and CH4 was also studied. This reaction was shown to be dependent on H4MPT, MFR, F420, H-S-CoM, and H-S-HTP.

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Abbreviations

AcCoA:

acetyl-CoA

AcP:

acetyl phosphate

H-S-CoM:

coenzyme M=2-mercaptoethanesulfonate

CH3-S-CoM:

methyl-coenzyme M=2-(methylthio)ethanesulfonate

H-S-HTP:

N-7-mercaptoheptanoylthreonine phosphate=component B

F420 :

coenzyme F420

H4MPT:

tetrahydromethanopterin

H4SPT:

tetrahydrosarcinapterin

MFR:

methanofuran

OH-B12 :

hydroxocobalamin

DTT:

dithiothreitol

MOPS:

morpholinopropane sulfonic acid

HPLC:

high performance liquid chromatography

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  1. Laboratorium für Mikrobiologie, Fachbereich Biologie, Philipps-Universität, Karl-von-Frisch-Strasse, D-3550, Marburg, Germany

    Reinhard Fischer & Rudolf K. Thauer

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  1. Reinhard Fischer
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  2. Rudolf K. Thauer
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Fischer, R., Thauer, R.K. Methyltetrahydromethanopterin as an intermediate in methanogenesis from acetate in Methanosarcina barkeri . Arch. Microbiol. 151, 459–465 (1989). https://doi.org/10.1007/BF00416607

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  • DOI: https://doi.org/10.1007/BF00416607

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