Optimizing E. coli as a formatotrophic platform for bioproduction via the reductive glycine pathway

Abstract

Microbial C1 fixation has a vast potential to support a sustainable circular economy. Hence, several biotechnologically important microorganisms have been recently engineered for fixing C1 substrates. However, reports about C1-based bioproduction with these organisms are scarce. Here, we describe the optimization of a previously engineered formatotrophic <i>Escherichia coli</i> strain. Short-term adaptive laboratory evolution enhanced biomass yield and accelerated growth of formatotrophic <i>E. coli</i> to 3.3 g-CDW/mol-formate and 6 h doubling time, respectively. Genome sequence analysis revealed that manipulation of acetate metabolism is the reason for better growth performance, verified by subsequent reverse engineering of the parental <i>E. coli</i> strain. Moreover, the improved strain is capable of growing to an OD₆₀₀ of 22 in bioreactor fed-batch experiments, highlighting its potential use for industrial bioprocesses. Finally, demonstrating the strain's potential to support a sustainable, formate-based bioeconomy, lactate production from formate was engineered. The optimized strain generated 1.2 mM lactate -10% of the theoretical maximum- providing the first proof-of-concept application of the reductive glycine pathway for bioproduction.