Monoethylene glycol (MEG) is an important commodity chemical with applications in numerous industrial processes， primarily in the manufacture of polyethylene terephthalate (PET) polyester used in packaging applications. In the drive towards a sustainable chemical industry， bio-based production of MEG from renewable biomass has attracted growing interest. Recent attempts for bio-based MEG production have investigated metabolic network modifications in Escherichia coli， specifically rewiring the xylose assimilation pathways for the synthesis of MEG. In the present study， we examined the suitability of Saccharomyces cerevisiae， a preferred organism for industrial applications， as platform for MEG biosynthesis. Based on combined genetic， biochemical and fermentation studies， we report evidence for the existence of an endogenous biosynthetic route for MEG production from D-xylose in S. cerevisiae which consists of phosphofructokinase and fructose-bisphosphate aldolase， the two key enzymes in the glycolytic pathway. Further metabolic engineering and process optimization yielded a strain capable of producing up to 4.0 g/L MEG， which is the highest titer reported in yeast to-date.