The rise in global temperature and accumulation of petroleum-based wastes in the environment forces the scientific focus towards renewable alternatives. In the present work, an under-exploited resource – spruce bark – is investigated as a raw material for production of bio-oil as a liquid energy carrier. To enhance the energy-content of the produced bio-crude, ultimately being produced through hydrothermal liquefaction, the polysaccharides were extracted through organosolv fractionation and converted to lipids by oleaginous microorganisms. The effect originating from tannins was also investigated by performing a pre-extraction before the organosolv fractionation. It was found that performing the organosolv fractionation and upgrading the isolated organosolv lignin to bio-oil greatly reduced the oxygen content of the oil fraction thereby improving its energy content, and introducing upgraded polysaccharides in the form of lipids, as well as pre-extracted tannins, caused clear changes in the product distribution of the final bio-oil and kept a final product with low oxygen content. The other factor largely influencing the product distribution originated from the various heating rates tested by altering operational mode of the HTL process between batch and semi-continuous. Ultimately, performing the organosolv fractionation and individual upgrading of the polysaccharides had a beneficial effect on reducing the final solids content and enhancing the liquid oil yield.