Enrichment of microbial helpers to enhance Methanosarcina growth and methane production using metagenomics and auxotrophy supplementation

This collection gather datasets and supplementary materials related to the following work.The main goal of this work was to investigate whether auxotrophies could be used to potentially reveal beneficial interactions between Methanosarcina and other species. To achieve this end, simplified anaerobic consortia were used to enrich microbial helpers that will in turn enhance Methanosarcina growth and methane production. Based on available metagenomic data from these simplified consortia (NCBI BioProject PRJNA1063026; Ginevra et al. 2025, Chemical Engineering Journal), the auxotrophies were predicted and several metagenome assembled genomes (MAGs) were selected as potentially beneficial for Methanosarcina spp ("helpers"). The predicted auxotrophies of the selected MAGs were used to supplement the simplified cultures with substrates, amino acids and/or vitamins in order to enrich the microbial helpers. The enriched cultures were followed during 3 months. Process data were monitored every 10 days (substrate consumption, gas production, etc.) and DNA samples were collected every 2 weeks to do metabarcoding (16S) and metagenomics analyses.