Rewiring methanol assimilation and reductive glycine pathways in Saccharomyces cerevisiae to increase one-carbon recovery

Abstract

Methanol is a promising substrate for biomanufacturing because of its low cost and non-food-competition. However, the bio-utilization of methanol presents significant challenges, including the requirement for glucose as a co-substrate, the toxicity of intermediate metabolite formaldehyde, and carbon loss during metabolism. These factors impede microbial growth and compromise the cost-effectiveness of the bioprocess. Here, we engineered Saccharomyces cerevisiae to grow on methanol while integrating the reductive glycine pathway (RGP) to reduce carbon loss. Our approach led to a 2.3-fold increase in methanol utilization and a 1.5-fold increase in biomass, coupled with CO2 reassimilation. The integration of methanol assimilation pathway with RGP mitigated formaldehyde toxicity and facilitated the co-utilization of CO2 and methanol. We provide the first evidence that CO2 and methanol can be co-utilized to synthesize pyruvate and mevalonate, key precursors of all terpenoids. As proof of principle, we complemented the engineered one-carbon utilization pathways in S. cerevisiae, achieving a 9.5-fold increase in cannabigerolic acid (CBGA). This study underscores the potential to broaden the spectrum of biosynthetic products derived from C1 compounds while facilitating carbon recovery. This research establishes a promising platform for sustainable biomanufacturing utilizing methanol and CO2.