Background: The bioconversion of cellulose into simple sugars or chemicals has attracted extensive attention in recent decades. The crystal allomorphs of cellulose are key factor affecting cellulose saccharification. However, due to the influence of lignin, hemicelluloses, and different characterization methods in the literature, the effect of cellulose allomorphs on cellulose saccharification is still unresolved. Thus, a systematic research on the effect of different cellulose allomorphs on enzymatic saccharification was required. Results: Multiple approaches, including the use of ionic liquid (IL), ethylenediamine (EDA), glycerol, and sodium hydroxide, were used to pretreat α-cellulose in this work. The properties of the obtained cellulose (crystallinity, lattice spacing, specific surface area, and wettability) were characterized by X-ray diffraction, Brunauer, Emmett, and Teller (BET) specific surface area analysis, and water contact angle analysis, respectively. The distance of the lattice spacing of cellulose III was longer than that of other cellulose samples. The crystallinity and water contact angles of the cellulose samples were ranked in the following order: cellulose treated with IL < cellulose treated with NaOH < cellulose treated with EDA < cellulose without treatment < cellulose treated with glycerol. Cellulose treated with IL, with a crystallinity index value of 20%, was very close to amorphous cellulose. After 72 h hydrolysis, the cellulose conversion ratio ranged from 43% to 99%. Cellulose treated with IL exhibited the best hydrolysis profile, followed by cellulose treated with EDA.