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The enzymatic hydrolysis of hardwood and softwood harvest residues treated by SO2-ethanol-water (SEW) fractionation was studied. The target was to convert these fibers with high yield into glucose monomers which could be further converted into biofuel by a subsequent fermentation stage. Hardwood biomass residues were efficiently digested at low enzyme dosage (5 FPU/g cellulose) whereas the softwood residues required notably higher enzyme dosage (20 FPU) for sufficient conversion. However, cellulase dosage of softwood could be reduced mannanase supplementation. Especially the high lignin content of softwood biomass pulps impairs the digestibility and thereby, improved delignification could notably enhance the hydrolysis yields. It was shown that inferior delignification of SW biomass is due to persistent polyphenolic acids present in coniferous bark, whereas no evidence of the negative effect of inorganics and acetone extractives was observed. Additionally, SW hydrolyzate was successfully converted into a mixture of butanol, acetone and ethanol through ABE fermentation. [ABSTRACT FROM AUTHOR]