To explore hydrolysis methods for the efficient manufacture of sugar solutions from the freshwater alga Water-net (Hydrodictyon reticulatum, HR), acid hydrolysis, enzymatic hydrolysis, and combined hydrolysis (acid followed by enzymatic hydrolysis) were investigated. In the one-step acid hydrolysis, the reaction of 8% solids content using 2% sulfuric acid at 120 o C for 1 hour was desirable. In this case, glucose 27.44 g 100 g DM-1 could be obtained from the HR-d13 samples. In the two-step acid hydrolysis, the primary hydrolysis (HR powder : 72% sulfuric acid = 1 g : 1.5 mL) was carried out for 1 hour at 60 o C, and then the secondary hydrolysis was done for 1 hour at 120 o C after addition of distilled water 23.5 mL. In this case, glucose 35.11 g/100 g DM could be obtained from the HR-d13 samples. In the combined hydrolysis, 25% solids content using 2% hydrochloric acid were reacted for 1 hour at 120 o C, and then citrate buffer and hydrolysis enzyme complexes (E1 1.0 mL+E2 0.2 mL g-1 dried matter) were added and reacted for 1-2 days at 50 o C. In this case, glucose 33.5 g 100 g DM-1 could be obtained from the HR-d23+26 samples. In conclusion, combined hydrolysis was likely to be more useful saccharification method of HR biomass at a practical level, considering the glucose productivity, generation of fermentation-inhibiting substances (hydroxyl methyl furfural, furfural), and limited use of strong acid.
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