D-xylose is a major component of lignocellulose and is after D-glucose the most abundant monosaccharide on earth. However, D-xylose cannot be naturally utilised by several industrially relevant microorganisms. On the way to a strong bio-based economy in Europe, this widely available feedstock has to be made accessible for the sustainable microbial synthesis of value-added chemical building blocks to be used in a broad range of applications. The project aims at engineering Corynebacterium glutamicum and Saccharomyces cerevisiae to serve as flexible and carbon efficient microbial cell factories converting D-xylose derived from lignocellulosic material (LCM) into value added products covering diols, lactols and organic acids. Combination of genetic engineering and systems biology are used to demonstrate the potential of a rational system-level approach for metabolic engineering of C. glutamicum and S. cerevisiae. In particular, new fundamental knowledge is generated regarding the impact of industrial hydrolysates on the intracellular dynamics of proteins, metabolites and fluxes as well as global stress responses and production capabilities of the two platform organisms. Subsequently, model-driven optimization approaches are expected to show the feasibility of growth-decoupled production with the newly constructed producer strains on industrial hydrolysates. The project has a high potential for innovative industrial applications, and it is expected to contribute to improve the competitiveness of the European biotechnological industry.