Abstract
Methanol represents a promising one-carbon substrate for high-value biochemicals and biofuels production. However, reprogramming a non-methylotrophic industrial host, such as Saccharomyces cerevisiae, to grow in minimal medium with methanol as sole carbon source is challenging. Here, a thermodynamic-based module-circuits (TMC) strategy followed by an adaptive laboratory evolution was adopted to engineer S. cerevisiae to efficiently utilize methanol in minimal medium. Through genome sequencing and reverse engineering, we found that the protein Shm2, involved in the glyoxylate-based serine (gSerine) pathway, plays a crucial role by reducing cytoplasmic accumulation of formaldehyde/formic acid. Through transcriptome and experimental validation, we identified that the ergosterol biosynthesis pathway was upregulated and the metabolites squalene and ergosterol were improved in our evolved strain, which were assumed to rescue cell in methanol medium by maintaining membrane integrity. Our results demonstrate that S. cerevisiae can be exploited as a promising one-carbon (C1) platform for biochemicals or biofuels production.
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