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Background  Formate converted from CO 2  reduction has great potential as a sustainable feedstock for biological production of biofuels and biochemicals. Nevertheless, utilization of formate for growth and chemical production by microbial species is limited due to its toxicity or the lack of a metabolic pathway. Here, we constructed a formate assimilation pathway in  Escherichia coli  and applied adaptive laboratory evolution to improve formate utilization as a carbon source in sugar-free conditions.    Results  The genes related to the tetrahydrofolate and serine cycles from  Methylobacterium extorquens  AM1 were overexpressed for formate assimilation, which was proved by the  13 C-labeling experiments. The amino acids detected by GC/MS showed significant carbon labeling due to biomass production from formate. Then, 150 serial subcultures were performed to screen for evolved strains with improved ability to utilize formate. The genomes of evolved mutants were sequenced and the mutations were associated with formate dehydrogenation, folate metabolism, and biofilm formation. Last, 90 mg/L of ethanol production from formate was achieved using fed-batch cultivation without addition of sugars.    Conclusion  This work demonstrates the effectiveness of the introduction of a formate assimilation pathway, combined with adaptive laboratory evolution, to achieve the utilization of formate as a carbon source. This study suggests that the constructed  E. coli  could serve as a strain to exploit formate and captured CO 2 .

Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions