The isolation of lignin is of great importance to understand its structural characteristics. A lithium chloride/dimethyl sulfoxide (LiCl/DMSO) solvent system has been developed for the dissolution of lignocellulose and for the isolation of lignin for this purpose. In this work, ball-milled wheat leaf (sheath included) was dissolved in the LiCl/DMSO solvent system and then regenerated in water. Two lignin preparations, cellulolytic enzyme lignin from the ball-milled leaf (CEL) and from the regenerated leaf (RCEL), were obtained through a cellulolytic enzyme lignin procedure. The RCEL and CEL were comparatively investigated by the use of wet chemistry and spectroscopic methods. The results indicate that the effects of ball milling and regeneration on the aromatic structure and β-O-4’ linkages of lignin were not significant. The RCEL had a higher isolation yield and purity, but a similar structure with the corresponding CEL. The RCEL can be used for structural analysis.