In this study, microcrystalline cellulose, which is a cell wall polysaccharide commonly contained in sea algae (brown algae, red algae and green algae), is used in substitution for cellulose and is hydrolyzed with seven enzymes available in the market. The seven enzymes selected are Viscozyme® L, Celluclast® 1.5 L, Saczyme, Novozym® 33095, Fungamyl® 800 L, Driselase® Basidiomycetes sp., and Alginate Lyase. To maximize the production of the reducing sugar by hydrolysis with each enzyme, we optimized the quantity of enzymes, reaction time, pH, and reaction temperature as four independent variables, and the reducing sugar production rate as a dependent variable, utilizing response surface methodology (RSM) to optimize the enzyme hydrolysis reaction conditions. Among the tested enzymes, the production rate of reducing sugar by Celluclast® 1.5 L was the highest. Hence, the predicted optimum conditions (8.5 % enzyme, reaction time 27.6 h, pH 4.1 and reaction temperature 44.1oC) were directly applied to Laminaria japonica and proved the predicted optimum conditions with experiments. Under the optimum conditions, the sugar yield of 137.6 mg/g-Laminaria japonica (experimental value) was obtained.