Engineering Yeast Yarrowia lipolytica for Methanol Assimilation

Conferring
methylotrophy on industrial microorganisms would enable
the production of diverse products from one-carbon feedstocks and
contribute to establishing a low-carbon society. Rebuilding methylotrophs,
however, requires a thorough metabolic refactoring and is highly challenging.
Only recently was synthetic methylotrophy achieved in model microorganismsEscherichia coli and baker’s yeast Saccharomyces cerevisiae. Here, we have engineered
industrially important yeast Yarrowia lipolytica to assimilate methanol. Through rationally constructing a chimeric
assimilation pathway, rewiring the native metabolism for improved
precursor supply, and laboratory evolution, we improved the methanol
assimilation from undetectable to a level of 1.1 g/L per 72 h and
enabled methanol-supported cellular maintenance. By transcriptomic
analysis, we further found that fine-tuning of methanol assimilation
and ribulose monophosphate/xylulose monophosphate (RuMP/XuMP) regeneration
and strengthening formate dehydrogenation and the serine pathway were
beneficial for methanol assimilation. This work paves the way for
creating synthetic methylotrophic yeast cell factories for low-carbon
economy.