Conversion of Escherichia coli to Generate All Biomass Carbon from CO2

Abstract

The living world is largely divided into autotrophs that convert CO₂ into biomass and heterotrophs that consume organic compounds. In spite of widespread interest in renewable energy storage and more sustainable food production, the engineering of industrially relevant heterotrophic model organisms to use CO₂ as their sole carbon source has so far remained an outstanding challenge. Here, we report the achievement of this transformation on laboratory timescales. We constructed and evolved Escherichia coli to produce all its biomass carbon from CO₂. Reducing power and energy, but not carbon, are supplied via the one-carbon molecule formate, which can be produced electrochemically. Rubisco and phosphoribulokinase were co-expressed with formate dehydrogenase to enable CO₂ fixation and reduction via the Calvin-Benson-Bassham cycle. Autotrophic growth was achieved following several months of continuous laboratory evolution in a chemostat under intensifying organic carbon limitation and confirmed via isotopic labeling.

Keywords: Escherichia coli; Rubisco; adaptive laboratory evolution; carbon fixation; metabolic rewiring; sustainability; synthetic autotrophy; synthetic biology.