AbstractA combination of steam explosion (SE) and alkaline peroxide (AP) used to pretreat bamboo was investigated. Steam explosion at 224 °C for 4 min was applied to bamboo, and the pretreated bamboo was delignified by alkaline peroxide. Enzymatic hydrolysis was compared in the pretreated samples. Steam pretreatment led to remarkable hemicellulose solubilization (63.2%). Lignin solubilization (93.1%) was achieved by alkaline peroxide treatment of steam-pretreated bamboo at 80 °C for 1 h in 0.88% (v/v) H2O2, whereas only 33.4% of lignin was solubilized when using raw bamboo. Pretreatment methods resulted in a low degree of polymerization and increased hydrolysis of cellulose. A maximum glucose yield of 90.5% was achieved with a combined steam explosion and alkaline peroxide pretreatment. The surface structure of treated bamboo and the adsorption of enzyme on the substrate were characterized by X-ray photoelectron spectroscopy. Delignification decreased enzyme adsorption and increased enzymatic conversion. SEM analyses indicated that SE-AP pretreatment disrupted lignin networks and exposed crystalline cellulose in bamboo more effectively than SE or AP pretreatment alone.