Owing to the low synthetic cost, low toxicity for most types and outstanding lignin solubility, deep eutectic solvents (DESs) have been recently exploited in fractionation of lignocellulosic biomass. Unfortunately, the DES-fractionated lignin, as another major product, was usually ignored and disposed improperly. Accordingly, the present study employed DES (choline chloride:lactic acid) to fractionate natural biomass for producing highly dispersed lignin nanoparticles. The obtained nano-lignin was utilized to prepare electrode materials for supercapacitors for the first time, and a higher specific capacitance than those prepared from conventional lignin was acquired. The results demonstrated that the lignin fractionation rate could be promoted via extending treatment time at low temperatures, whereas the DES to biomass mass ratio might be the lignin dissolution-limiting factor at high temperature. Uniform lignin nanoclusters could be obtained after DES fractionation, and increasing fractionation temperature was beneficial for preparing nano-lignin in DES. The cleavages of lignin-carbohydrate complexes and lignin subunits might account for the formation mechanism of lignin oligomers. Meanwhile, the aggregation of monolignols could contribute to the growth of lignin nanoparticles. The one-step KOH activation could induce hierarchical pores in lignin fractionated from moderate residence time, whereas two-step method was advisable for more aggregated DES-lignin. Based on the directly activated DES-lignin with a high surface area of 3577.3 m
2
g
-1
, the prepared supercapacitor exhibited a superior specific capacitance of 248.8 F g
-1
. This study aimed to provide a novel method for producing high-performance energy storage carbon materials based on DES fractionated lignin from lignocellulose.
