Softwood residues are the most abundant feedstock available for bioconversion in many northern countries. However, the high costs for delignification and enzymatic hydrolysis currently deter commercialization of softwood bioconversion processes. This study evaluates the abilities of two novel fungal preparations (MSUBC1 and MSUBC2) and two commercial cellulase preparations (TR1 and TR2) to hydrolyze cellulose in Douglas-fir pretreated by steam explosion or ethanol organosolv process. MSUBC1 AA showed significantly better performance than the other preparations on both lignocellulosic substrates. In particular, MSUBC1 achieved > 76% cellulose conversion for hydrolysis of steam-exploded Douglas-fir (similar to 44% lignin) after 72 h at low enzyme loading (10 filter paper units/g of cellulose) and without beta-glucosidase supplementation.