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Grass clipping, a cellulose-rich raw material, has great potential to produce biofuels, but must be firstly hydrolyzed to liberate fermentable sugars. In this study, grass clipping was pretreated with ultrasound (US), Ca (OH) 2 , NaOH, US-Ca(OH) 2 and US-NaOH at relatively low temperature to enhance its enzymatic hydrolysis. The solubilization of hemicellulose and lignin, and crystallinity index of cellulose increased after US-alkaline pretreatment, leading to a significant increase of enzyme accessibility to cellulose. Compared with another four pretreatments, US-Ca(OH) 2 pretreatment of grass clipping showed the best improvement for reducing sugar yield. X-ray diffraction (XRD) determination and scanning electron microscope (SEM) observation showed that the crystallinity index of grass clipping increased and the grass clipping surface suffered from serious erosion after US-Ca(OH) 2 pretreatment. Then, the operating conditions of US-Ca(OH) 2 pretreatment and enzymatic hydrolysis were systematically optimized, and the suitable operating conditions were as follows: US power density of 0.65 W/ml, US pretreatment time of 30 min, Ca(OH) 2 concentration of 0.75%, pretreatment temperature of 75°C, enzyme loading of 125 FPU/g, and hydrolysis time of 72 h. The reducing sugar yield of grass clipping pretreated by US-Ca(OH) 2 reached 414 mg/g, increasing by 3.5 times compared with that of raw grass clipping. The US-Ca(OH) 2 pretreatment of grass clipping at low temperature significantly enhanced the potential of grass clipping as a promising raw material to produce biofuels.

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