A unified and simple medium for growing model methanogens

Apart from their archetypic use in anaerobic digestion (AD) methanogenic archaea are targeted for a wide range of applications. Using different methanogenic archaea for one specific application requires the optimization of culture media to enable the growth of different strains under identical environmental conditions, e.g., in microbial electrochemical technologies (MET) for (bio)electromethanation. Here we present a new culture medium (BFS01) adapted from the DSM-120 medium by omitting resazurin, yeast extract, casitone, and using a low salt concentration, that was optimized for Methanosarcina barkeri, Methanobacterium formicicum, and Methanothrix soehngenii. The aim was to provide a medium for follow-up co-culture studies using specific methanogens and Geobacterspp. dominated biofilm anodes. All three methanogens showed growth and activity in the BFS01 medium. This was demonstrated by estimating the specific growth rates (mu) and doubling times (t(d)) of each methanogen. The mu and t(d) based on methane accumulation in the headspace showed values consistent with literature values for M. barkeri and M. soehngenii. However, mu and t(d) based on methane accumulation in the headspace differed from literature data for M. formicicum but still allowed sufficient growth. The lowered salt concentration and the omission of chemically complex organic components in the medium may have led to the observed deviation from mu and t(d) for M. formicicum as well as the changed morphology. 16S rRNA gene-based amplicon sequencing and whole genome nanopore sequencing further confirmed purity and species identity.

Automated summary

By optimizing the culture medium, this study successfully achieved growth and activity in Methanosarcina barkeri, Methanobacterium formicicum, and Methanothrix soehngenii. This finding provides a foundation for follow-up co-culture studies and demonstrates that the optimized culture medium can effectively support the growth of these methanogens.