Lithium-sulfur (Li-S) batteries have arousing interest because of their high theoretical energy density. However, they often suffer from sluggish conversion of lithium polysulfides (LiPS) during the charge/discharge process. Single nickel (Ni) atoms on nitrogen-doped graphene (Ni@NG) with Ni-N structure are prepared and introduced to modify the separators of Li-S batteries. The oxidized Ni sites of the Ni-N structure act as polysulfide traps, efficiently accommodating polysulfide ion electrons by forming strong S ⋅⋅⋅NiN bonding. Additionally, charge transfer between the LiPS and oxidized Ni sites endows the LiPS on Ni@NG with low free energy and decomposition energy barrier in an electrochemical process, accelerating the kinetic conversion of LiPS during the charge/discharge process. Furthermore, the large binding energy of LiPS on Ni@NG also shows its ability to immobilize the LiPS and further suppresses the undesirable shuttle effect. Therefore, a Li-S battery based on a Ni@NG modified separator exhibits excellent rate performance and stable cycling life with only 0.06% capacity decay per cycle. It affords fresh insights for developing single-atom catalysts to accelerate the kinetic conversion of LiPS for highly stable Li-S batteries.