DOI: 10.5061/dryad.931zcrjvd
title: Nature and nurture: Enabling formate-dependent growth in Methanosarcina acetivorans

description: Methanogenic archaea are crucial in global carbon cycling as they produce
 around 1 Gt of the potent greenhouse gas methane annually. Major
 contributors belong to the order Methanosarcinales, represented by
 versatile methanogens that are capable of acetoclastic, methylotrophic,
 and CO2-reducing methanogenesis. The genetically tractable model
 methanogen Methanosarcina acetivorans by its nature shows versatility in
 substrate utilization and energy conservation pathways but cannot utilize
 formate. In this study, we expanded the primary metabolism of M.
 acetivorans to include formate-dependent methanogenesis. By introducing
 formate dehydrogenase, two metabolically engineered M. acetivorans strains
 acquired the capacity for formate-dependent methanogenesis. One catabolic
 pathway relies on formate-dependent methyl-reduction, and the other relies
 on formate-dependent CO2-reduction. Nurturing the M. acetivorans strain
 capable of CO2-reduction with adaptive laboratory evolution enabled growth
 and methanogenesis solely on formate, a metabolism only reported in
 methanogens without cytochromes. M. acetivorans also showed acetogenic
 potential where the formate-dependent CO2-reducing strain was able to
 divert ≈ 10% of carbon from the substrates provided to acetate instead of
 methane. Even though M. acetivorans lacks energy-converting hydrogenase
 and cannot use H2, we show that M. acetivorans has a yet-uncharacterized
 capacity to obtain reduced ferredoxins from oxidizing formate. Our work
 encourages a reevaluation of our understanding of formate utilization in
 Methanosarcinales. By enabling formate-dependent methanogenesis, we have
 expanded the substrate spectrum of a versatile model methanogen with
 cytochromes to include formate.
description: # Nature and nurture: Enabling formate-dependent growth in *Methanosarcina
 acetivorans*
 [https://doi.org/10.5061/dryad.931zcrjvd](https://doi.org/10.5061/dryad.931zcrjvd) The dataset contains the raw data obtained to plot the growth curves (OD600 values) and the metabolite curves (Methane as measured using gas chromatography and methanol, formate, acetate, and pyruvate as measured using NMR). It also contains information on the media composition used in the experiments. ## Description of the data and file structure The Excel file contains 5 sheets- WWM73, JB-MF, JB-F2_HSF, JB-F2_HSMe, Media Each Excel sheet contains the first column as time points and the rest of the columns in the format: media-strain-metabolite-replicate number For example in Sheet WWM73--- The heading of Column 2 is MF-WWM73-OD-1 where MF refers to the media in which strain WWM73 was grown. The data obtained is of optical density values of the first replicate. Sheet WWM73 contains raw data for the Figure 4. Cells with 'n/a' represent data not available as NMR samples were not measured for the given time points. Sheet JB-MF contains raw data for the Figure 6. Sheets JB-F2_HSF and JB-F2_HSMe contain raw data for Figure 7. Cells with 'n/a' represent data not available as NMR samples were not measured for the given time points. Sheet Media contains an explanation of the media abbreviation and composition. The first column is time (hours) ## Access information NA