Abstract

Methanol is generally metabolized through a pathway initiated by a cobalamine-containing methanol methyltransferase by anaerobic methylotrophs (such as methanogens and acetogens), or through oxidation to formaldehyde using a methanol dehydrogenase by aerobes. Methanol is an important substrate in deep-subsurface environments, where thermophilic sulfate-reducing bacteria of the genus Desulfotomaculum have key roles. Here, we study the methanol metabolism of Desulfotomaculum kuznetsovii strain 17T, isolated from a 3000-m deep geothermal water reservoir. We use proteomics to analyze cells grown with methanol and sulfate in the presence and absence of cobalt and vitamin B12. The results indicate the presence of two methanol-degrading pathways in D. kuznetsovii, a cobalt-dependent methanol methyltransferase and a cobalt-independent methanol dehydrogenase, which is further confirmed by stable isotope fractionation. This is the first report of a microorganism utilizing two distinct methanol conversion pathways. We hypothesize that this gives D. kuznetsovii a competitive advantage in its natural environment.

Highlights

  • Methanol is generally metabolized through a pathway initiated by a cobalamine-containing methanol methyltransferase by anaerobic methylotrophs, or through oxidation to formaldehyde using a methanol dehydrogenase by aerobes

  • The presence of genes coding for a methanol MT system in the genome of D. kuznetsovii suggested the involvement of a vitamin B12-dependent MT system in methanol conversion[7], while previous analysis indicated the involvement of an alcohol dehydrogenase[34]

  • When cobalt and vitamin B12 were omitted from the medium D. kuznetsovii was still able to degrade methanol, but the residual methanol concentration at the end of the assays was significantly higher (p = 0.00027) than in assays with cobalt and vitamin B12 (Supplementary Fig. 1)

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Summary

Introduction

Methanol is generally metabolized through a pathway initiated by a cobalamine-containing methanol methyltransferase by anaerobic methylotrophs (such as methanogens and acetogens), or through oxidation to formaldehyde using a methanol dehydrogenase by aerobes. Growth of D. kuznetsovii with methanol and sulfate was studied and resulted in a partially purified alcohol dehydrogenase (ADH) with a molecular mass of 42 kDa that showed activity with methanol, but activity with ethanol was ten times higher[34].

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