A novel natural deep eutectic solvent (NDES) with water content ranging from 65 to 93 wt%, in which betaine (Bet) acts as the cation and amino acids (AAs) as the anions, was prepared by a simple and green chemical route. [Bet][AA] NDES showed excellent xylan and lignin solubility, however, scare cellulose solubility. A mild and facile pretreatment process with [Bet][AA] NDES was carried out at 60 °C for 5 h. The enzymatic hydrolysis efficiency of cellulose and corncob was significantly improved. Detailed characterization showed that the enhancement of cellulose digestibility derived mainly from xylan and lignin removal. Xylan and lignin removal for [Bet][Lys]-W87 was 47.68 and 49.06%, while it was 42.20% and 57.01% for [Bet][Arg]-W82, respectively. FT-IR, SEM, XRD, and HSQC NMR studies confirmed the effectiveness and mechanism of [Bet][Lys]-W87 and [Bet][Arg]-W82 on biomass pretreatment.