We used the eQuilibrator API to calculate each compound's standard free energy of formation, ΔG f '°, and the change in free energy for each reaction, ΔG r '°6 , estimated at standard conditions, meaning that all species are at 1 M concentration at 25°C and 1 bar (see Figure ). We used the eQuilibrator API default value for the Mg 2+ concentration (pMg, in this case pMg = 3.0), which is required for these calculations, but which mostly affects reactions involving ATP. We explored the eQuilibrator API calculations at pH 10. The eQuilibrator software uses component contribution methods to estimate thermodynamic values. Temperature and pressure cannot be varied, because the model calibration relies on experimental measurements for ΔfG conducted at standard T and P. The Neo4J defined search pattern doesn't guarantee a pathway identified as topologically autocatalytic is energetically favorable, since the reactions derived from reaction expansion do not specifically encode energetic information. To address this problem, we merged computed thermochemical information derived from the eQuilibrator API onto the CRNR nodes so that network queries could constrain energetic favorability. For example, sorting the pattern match results by the minimum aggregated energy across the reactions in the ring path and returning the lowest energy paths should yield the most energetically favorable reaction motifs.