Constructing artificial methylotrophic Saccharomyces cerevisiae for chondroitin sulfate production using a multidimensional combinatorial strategy
The concept of a methanol-based circular bioeconomy envisions the conversion of greenhouse gases into one-carbon compounds, which are subsequently utilized to produce high-value-added products. However, the toxicity of intermediates limits the metabolic efficiency within chassis strains. Here, a carbon metabolism reconfiguration strategy combined with adaptive laboratory evolution (ALE) was applied to engineer an artificial methylotrophic yeast by accelerating the flux of toxic intermediates into the central carbon metabolism. After modifying the key gene Sfa1 identified by ALE and introducing a light-responsive switch to dynamically redirect the carbon flux, the engineered strain not only restored its growth in minimal medium with 2% methanol, but also synthesized chondroitin sulfate A driven by the dark reaction. Furthermore, we designed a 3'-phosphoadenosine-5'-phosphosulfate supply strategy and energy adapter to increase the sulfonation level. This study illustrates the exciting potential of methylotrophic S. cerevisiae as a platform for biosynthesis.