Diffusion of the Interspecies Electron Carriers H ₂ and Formate in Methanogenic Ecosystems and Its Implications in the Measurement of <i>K</i> _<i>m</i> for H ₂ or Formate Uptake

We calculated the potential H 2 and formate diffusion between microbes and found that at H 2 concentrations commonly found in nature, H 2 could not diffuse rapidly enough to dispersed methanogenic cells to account for the rate of methane synthesis but formate could. Our calculations were based on individual organisms dispersed in the medium, as supported by microscopic observations of butyrate-degrading cocultures. We isolated an axenic culture of Syntrophomonas wolfei and cultivated it on butyrate in syntrophic coculture with Methanobacterium formicicum ; during growth the H 2 concentration was 63 nM (10.6 Pa). S. wolfei contained formate dehydrogenase activity (as does M. formicicum ), which would allow interspecies formate transfer in that coculture. Thus, interspecies formate transfer may be the predominant mechanism of syntrophy. Our diffusion calculations also indicated that H 2 concentration at the cell surface of H 2 -consuming organisms was low but increased to approximately the bulk-fluid concentration at a distance of about 10 μm from the surface. Thus, routine estimation of kinetic parameters would greatly overestimate the K m for H 2 or formate.