Rice straw has recently attracted interest in Japan as a potential source of raw material for ethanol production. Wet disk milling, a continuous pretreatment to enhance the enzymatic digestibility of rice straw, was compared with conventional ball milling and hot-compressed water treatment. Pretreated rice straw was evaluated by enzymatic hydrolysis using Acremonium cellulase and characterized by X-ray diffraction and scanning electron microscopy. Glucose and xylose yields by wet disk milling, ball milling, and hot-compressed water treatment were 78.5% and 41.5%, 89.4% and 54.3%, and 70.3% and 88.6%, respectively. Wet disk milling and hot-compressed water treatment increased sugar yields without decreasing their crystallinity. The feature size of the wet disk milled rice straw was similar to that of hot-compressed water-treated rice straw. The energy consumption of wet disk milling was lower than that of other pretreatments. Thus, wet disk milling is an economical, practical pretreatment for the enzymatic hydrolysis of lignocellulosic biomass, especially herbaceous biomass such as rice straw. © 2009 Elsevier Ltd. All rights reserved.