Highlights • Conversion of obligate heterotroph to full autotrophy over laboratory timescales • Non-native Calvin cycle operation generates biomass carbon from CO2 in E. coli • Formate is oxidized by heterologous formate dehydrogenase to provide reducing power • Chemostat-based directed evolution led to complete trophic mode change in ≈200 days Summary The living world is largely divided into autotrophs that convert CO2 into biomass and heterotrophs that consume organic compounds.
