Hot-compressed water (HCW) treatment without chemicals such as acids, bases or organic solvents is an environmentally-benign pretreatment method to enhance the enzymatic digestibility of cellulose and hemicellulose components in the lignocellulosic biomass. An enhanced enzymatic digestibility of the HCW-treated cellulose residue is related to the removal rate of hemicellulose and lignin in the feedstock. Meanwhile, the sugars from the hemicellulose fraction are largely decomposed to other compounds such as furfurals in a severe HCW condition that enhances the enzymatic digestibility of cellulose residue. To improve total sugar yields from both components, we investigated the effects of a two-step HCW treatment on the enzymatic hydrolysis of eucalyptus (hardwood) and Douglas-fir (softwood). The optimum conditions of the first HCW treatment for hemicellulose solubilization of eucalyptus and Douglas-fir were at 210˚C for 5 min and at 220˚C for 5 min, respectively. Each of the resultant residues was washed and then treated by the second HCW treatment at 240-260˚C for 5 min to increase enzymatic digestibility of cellulose. The eucalyptus treated with two-step HCW (210˚C/240˚C) was converted to an approximately 490 g of total sugar/kg-wood with a cellulase loading of 10 FPU/g-residue. The Douglas-fir treated with two-step HCW (220˚C/260˚C) was converted to an approximately 440 g of total sugar/kg-wood, though the high cellulase loading of 40 FPU/g-residue was required for enzymatic hydrolysis of the residue. The total sugar yields from these woody biomass treated by two-step HCW were 1.3-1.5 times higher than those by the optimized one-step HCW.