Lignocellulosic biomass pretreatment with deep eutectic solvents (DESs) is a promising and challenging process for production of biofuels and valuable platform chemicals. In this work, rice straw was mainly fractionated into carbohydrate-rich materials (CRMs) and lignin-rich materials (LRMs) by 90% lactic acid/choline chloride (LC)-water solution with different molar ratio of hydrogen bond donor (HBD, lactic acid) and hydrogen bond acceptor (HBA, choline chloride). It was found that high HBD/HBA molar ratio of DESs was favorable for achieving CRMs and LRMs with high purity, and both HBD and HBA were responsible for effective biomass fractionation possibly due to their synergistic effect on highly efficient breakage of the linkage between hemicellulose and lignin and thus lignin extraction. About 30%-35% of lignin in native rice straw was fractionated as LRMs, and exceeding 70% of xylan were removed and fractionated into the liquid stream as forms of xylose, furfural and humins after pretreatment using aqueous LC (3:1, 5:1) solution. Consequently, polysaccharides enzymatic hydrolysis of the CRMs were significantly enhanced. Moreover, all the DESs could be recovered with high yields of around 90%, and 69% of the LC (3:1) was recovered after 5 cycles reuse at 90 degrees C. Besides, the recycled DES maintained a good pretreatment ability, and glucose yields of 60-70% were achieved in the enzymatic hydrolysis of CRMs obtained in each cycle. The facile process established in present work is promising for large scale production of fermentable sugars and other chemicals. (C) 2018, The Society for Biotechnology, Japan. All rights reserved.