Conversion of Escherichia coli into Mixotrophic CO₂ Assimilation with Malate and Hydrogen Based on Recombinant...

We report the mixotrophic growth of Escherichia coli based on recombinant 2-oxoglutarate:ferredoxin oxidoreductase (OGOR) to assimilate CO₂ using malate as an auxiliary carbon source and hydrogen as an energy source. We employ a long-term (~184 days) two-stage adaptive evolution to convert heterotrophic E. coli into mixotrophic E. coli. In the first stage of evolution with serine, diauxic growth emerges as a prominent feature. At the end of the second stage of evolution with malate, the strain exhibits mixotrophy with CO₂ as an essential substrate for growth. We expect this work will open new possibilities in the utilization of OGOR for microbial CO₂ assimilation and future hydrogen-based electro-microbial conversion.